Inhibitors of the ICE/ced-3 family of cysteine proteases

ABSTRACT

This invention is directed to novel sulfonimide (substituted)acyl dipeptidyl ICE/ced-3 family inhibitor compounds having the following structure:                    
     wherein A, B, X, R, R 1 , R 2 , n, q, and r are as defined herein. The invention is also directed to pharmaceutical compositions containing these compounds, as well as the use of such compositions in the treatment of patients suffering inflammatory, autoimmune and neurodegenerative diseases, for the prevention of ischemic injury, and for the preservation of organs that are to undergo a transplantation procedure.

CROSS REFERENCE TO RELATED APPLICATION

This application claims priority from U.S. Provisional Application No.60/327,555 filed Apr. 17, 2000, which provisional application wasoriginally filed as U.S. application Ser. No. 09/550,917 on Apr. 17,2000 and converted to the above provisional application by Petitionfiled Apr. 17, 2001.

TECHNICAL FIELD

The present invention relates to novel classes of compounds which areinhibitors of interleukin-1β converting enzyme and related proteases(“ICE/ced-3 family of cysteine proteases”), as well as to pharmaceuticalcompositions comprising these compounds and to methods of using suchpharmaceutical compositions.

BACKGROUND OF THE INVENTION

Interleukin 1 (“IL-1”) is a major pro-inflammatory and immunoregulatoryprotein that stimulates fibroblast differentiation and proliferation,the production of prostaglandins, collagenase and phospholipase bysynovial cells and chondrocytes, basophil and eosinophil degranulationand neutrophil activation. Oppenheim, J. H. et al., Immunology Today7:45-56 (1986). As such, it is involved in the pathogenesis of chronicand acute inflammatory and autoimmune diseases. IL-1 is predominantlyproduced by peripheral blood monocytes as part of the inflammatoryresponse. Mosely, B. S. et al., Proc. Nat. Acad. Sci. 84:4572-4576(1987); Lonnemann, G. et al., Eur. J. Immunol. 19:1531-1536 (1989).

IL-1β is synthesized as a biologically inactive precursor, proIL-1β.ProIL-1β is cleaved by a cysteine protease called interleukin-1βconverting enzyme (“ICE”) between Asp-116 and Ala-117 to produce thebiologically active C-terminal fragment found in human serum andsynovial fluid. Sleath, P. R. et al., J. Biol. Chem. 265:14526-14528(1992); A. D. Howard et al., J. Immunol. 147:2964-2969 (1991).

ICE is a cysteine protease localized primarily in monocytes. In additionto promoting the pro-inflammatory and immunoregulatory properties ofIL-1β, ICE, and particularly its homologues, also appear to be involvedin the regulation of cell death or apoptosis. Yuan, J. et al., Cell75:641-652 (1993); Miura, M. et al., Cell 75:653-660 (1993);Nett-Giordalisi, M. A. et al., J. Cell Biochem. 17B:117 (1993). Inparticular, ICE or ICE/ced-3 homologues are thought to be associatedwith the regulation of apoptosis in neurogenerative diseases, such asAlzheimer's and Parkinson's disease. Marx, J. and M. Baringa, Science259:760-762 (1993); Gagliardini, V. et al., Science 263:826-828 (1994).

Thus, disease states in which inhibitors of the ICE/ced-3 family ofcysteine proteases may be useful as therapeutic agents include:infectious diseases, such as meningitis and salpingitis; septic shock,respiratory diseases; inflammatory conditions, such as arthritis,cholangitis, colitis, encephalitis, endocerolitis, hepatitis,pancreatitis and reperfusion injury, ischemic diseases such as themyocardial infarction, stroke and ischemic kidney disease; immune-baseddiseases, such as hypersensitivity; auto-immune diseases, such asmultiple sclerosis; bone diseases; and certain neurodegenerativediseases, such as Alzheimer's and Parkinson's disease. Such inhibitorsare also useful for the repopulation of hematopoietic cells followingchemo- and radiation therapy and for prolonging organ viability for usein transplantation.

ICE/ced-3 inhibitors represent a class of compounds useful for thecontrol of the above-listed disease states. Peptide and peptidylinhibitors of ICE have been described. However, such inhibitors havebeen typically characterized by undesirable pharmacologic properties,such as poor oral absorption, poor stability and rapid metabolism.Plattner, J. J. and D. W. Norbeck, in Drug Discovery Technologies, C. R.Clark and W. H. Moos, Eds. (Ellis Horwood, Chichester, England, 1990),pp. 92-126. These undesirable properties have hampered their developmentinto effective drugs.

Accordingly, the need exists for compounds that can effectively inhibitthe action of the ICE/ced-3 family of proteases, for use as agents forpreventing unwanted apoptosis and for treating chronic and acute fonnsof IL-1 mediated diseases, such as inflammatory, autoimmune orneurodegenerative diseases. The present invention satisfies this needand provides further related advantages.

SUMMARY OF THE INVENTION

In general, the compounds of this invention incorporate an aryl orheteroaryl substituted acyl group as a dipeptide mimetic. The resultingcompounds exhibit improved properties relative to their peptidiccounterparts, for example, such as improved cell penetration or improvedabsorption and metabolic stability resulting in enhancedbioavailability.

One aspect of the instant invention is the compounds of the Formula I:

wherein A, B, X, n, q, r, R, R¹ and R² are as defined below, as well aspharmaceutically acceptable salts thereof.

A further aspect of the instant invention is a pharmaceuticalcomposition comprising a compound of the above Formula I and apharmaceutically-acceptable carrier therefor.

Another aspect of this invention involves a method for treating anautoimmune disease comprising administering an effective amount of apharmaceutical composition discussed above to a patient in need of suchtreatment.

Yet another aspect of the instant invention is a method for treating aninflammatory disease comprising administering an effective amount of apharmaceutical composition discussed above to a patient in need of suchtreatment.

A further aspect of the instant invention is a method for treating aneurodegenerative disease comprising administering an effective amountof a pharmaceutical composition discussed above to a patient in need ofsuch treatment.

Another aspect of the instant invention is a method of preventingischemic injury to a patient suffering from a disease associated withischemic injury comprising administering an effective amount of thepharmaceutical composition discussed above to a patient in need of suchtreatment.

A further aspect of the instant invention is a method for expanding ofhematopoietic cell populations and/or enhancing their survival bycontacting the cells with an effective amount of the pharmaceuticalcomposition discussed above. Cell populations included in the method ofthe invention include (but are not limited to) granulocytes, monocytes,erthrocytes, lymphocytes and platelets for use in cell transfusions.

An alternate aspect of the instant invention is a method of prolongingthe viability of an organ that has been removed from the donor for thepurpose of a future transplantation procedure, which comprises applyingan effective amount of the pharmaceutical composition discussed above tothe organ, thereby prolonging the viability of the organ as compared toan untreated organ. The organ may be an intact organ, or isolated cellsderived from an organ (e.g., isolated pancreatic islet cells, isolateddopaminergic neurons, blood or hematopoietic cells).

These and other aspects of this invention will be evident upon referenceto the following detailed description.

DETAILED DESCRIPTION OF THE INVENTION

As mentioned above, one aspect of the instant invention is the compoundsof the Formula I:

wherein:

n is 0, 1 or 2;

q is 1 or 2;

r is 1 or 2;

R is lower alkyl, alkyl, cycloalkyl, (cycloalkyl)alkyl, phenyl,substituted phenyl, phenylalkyl, substituted phenylalkyl, naphthyl,substituted naphthyl, (1 or 2 naphthyl)alkyl, substituted (1 or 2naphthyl)alkyl, heteroaryl, substituted heteroaryl, (heteroaryl)alkyl,substituted (heteroaryl)alkyl, NR^(a)(R^(b)) or OR^(c);

R¹ is phenyl, substituted phenyl, naphthyl, substituted naphthyl,heteroaryl, or substituted heteroaryl;

R² is hydrogen, alkyl, cycloalkyl, phenyl, substituted phenyl,(CH₂)_(p)CO₂R³, (CH₂)_(m)NH₂, (CH₂)_(m)NHCOR¹⁰, (CH₂)_(m)N(C═NH)NH₂,(CH₂)_(p)OR¹¹, (CH₂)_(p)SR¹², (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or 2-naphthyl), or(CH₂)_(m)heteroaryl, wherein heteroaryl includes (but is not limited to)substituted or unsubstituted pyridyl, thienyl, furyl, thiazolyl,imidazolyl, pyrazolyl, isoxazolyl, pyrazinyl, pyrimidyl, triazinyl,tetrazolyl, and indolyl;

X is CH₂, C═O, O, S, NH, C═ONH or CH₂OC═ONH;

A is a natural or unnatural amino acid of Formula IIa-i:

 B is a hydrogen atom, a deuterium atom, C₁₋₁₀ straight chain orbranched alkyl, cycloalkyl, phenyl, substituted phenyl, naphthyl,substituted naphthyl, 2-benzoxazolyl, substituted 2-oxazolyl,(CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl),(CH₂)_(m)(1 or 2-naphthyl), (CH₂)_(m)heteroaryl, halomethyl, CO₂R¹³,CONR¹⁴R¹⁵, CH₂ZR¹⁶, CH₂OCO(aryl), CH₂OCO(substituted aryl),CH₂OCO(heteroaryl), CH₂OCO(substituted heteroaryl), or CH₂OPO(R¹⁷)R¹⁸,where Z is an oxygen or a sulfur atom, or B is a group of the FormulaIIIa-c:

 and wherein

R^(a) and R^(b) are the same or different and independently hydrogen,alkyl, cycloalkyl, (cycloalkyl)alkyl, phenyl, substituted phenyl,phenylalkyl, substituted phenylalkyl, naphthyl, substituted naphthyl, (1or 2 naphthyl)alkyl, substituted (1 or 2 naphthyl)alkyl, heteroaryl,substituted heteroaryl, (heteroaryl)alkyl, or substituted(heteroaryl)alkyl, with the proviso that R^(a) and R^(b) cannot both behydrogen;

R^(c) is alkyl, cycloalkyl, (cycloalkyl)alkyl, phenyl, substitutedphenyl, phenylalkyl, substituted phenylalkyl, naphthyl, substitutednaphthyl, (1 or 2 naphthyl)alkyl, substituted (1 or 2 naphthyl)alkyl,heteroaryl, substituted heteroaryl, (heteroaryl)alkyl, or substituted(heteroaryl)alkyl;

R³ is hydrogen, alkyl, cycloalkyl, (cycloalkyl)alkyl, phenylalkyl, orsubstituted phenylalkyl;

R⁴ is alkyl, cycloalkyl, phenyl, substituted phenyl, (CH₂)_(m)NH₂,(CH₂)_(m)NHCOR¹⁰, (CH₂)_(m)N(C═NH)NH₂, (CH₂)_(p)CO₂R³, (CH₂)_(p)OR¹¹,(CH₂)_(p)SR¹², (CH₂)_(m)Cycloalkyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or 2-naphthyl), or(CH₂)_(m)heteroaryl, wherein heteroaryl includes (but is not limited to)pyridyl, thienyl, furyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl,pyrazinyl, pyrimidyl, triazinyl, tetrazolyl, and indolyl,

R^(4a) is hydrogen or methyl, or

R⁴ and R^(4a) taken together are —(CH₂)_(d)— where d is an integer from2 to 6;

R⁵ is phenyl, substituted phenyl, (CH₂)_(p)phenyl, (CH₂)_(p)(substitutedphenyl), cycloalkyl, or benzofused cycloalkyl;

R⁶ is hydrogen, alkyl, cycloalkyl, phenyl, substituted phenyl,(CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), or(CH₂)_(m)(1 or 2-naphthyl);

R⁷ is hydrogen, fluorine, oxo (i.e., ═O), alkyl, cycloalkyl, phenyl,substituted phenyl, naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or 2-naphthyl), OR¹¹, SR¹²,or NHCOR¹⁰;

R⁸ is hydrogen, oxo, alkyl, cycloalkyl, phenyl, substituted phenyl,naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substitutedphenyl), or (CH₂)_(m)(1 or 2-naphthyl);

R⁹ is alkyl, cycloalkyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or 2-naphthyl), or COR¹⁰;

R¹⁰ is hydrogen, alkyl, cycloalkyl, phenyl, substituted phenyl,naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substitutedphenyl), (CH₂)_(m)(1 or 2-naphthyl), OR¹³, or NR¹⁴R¹⁵;

R¹¹ is hydrogen, alkyl, cycloalkyl, phenyl, substituted phenyl,naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substitutedphenyl), or (CH₂)_(m)(1 or 2-naphthyl);

R¹² is alkyl, cycloalkyl, phenyl, substituted phenyl, naphthyl,(CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), or(CH₂)_(m)(1 or 2-naphthyl);

R¹³ is alkyl, cycloalkyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), or (CH₂)_(m)(1 or 2-naphthyl);

R¹⁴ is hydrogen, alkyl, cycloalkyl, phenyl, substituted phenyl,naphthyl, substituted naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), or (CH₂)_(m)(1 or 2-naphthyl);

R¹⁵ is hydrogen or alkyl; or

R¹⁴ and R¹⁵ taken together form a five, six or seven memberedcarbocyclic or heterocyclic ring, such as morpholine or N-substitutedpiperazine;

R¹⁶ is phenyl, substituted phenyl, naphthyl, substituted naphthyl,heteroaryl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1or 2-naphthyl), or (CH₂)_(m)heteroaryl;

R¹⁷ and R¹⁸ are independently alkyl, cycloalkyl, phenyl, substitutedphenyl, naphthyl, or phenylalkyl, substituted phenylalkyl, or(cycloalkyl)alkyl;

R¹⁹ and R²⁰ are independently hydrogen, alkyl, phenyl, substitutedphenyl, (CH₂)_(m)phenyl, or (CH₂)_(m)(substituted phenyl), or R¹⁹ andR²⁰ taken together are —(CH═CH)₂—;

R²¹ hydrogen, alkyl, phenyl, substituted phenyl, (CH₂)_(m),phenyl,(CH₂)_(m)(substituted phenyl);

R²², R²³ and R²⁴ are independently hydrogen or alkyl;

Y¹ is CH₂, (CH₂)₂, (CH₂)₃, or S;

Y² is O or NR²⁴;

Y³ is CH₂, O, or NR²⁴;

a is 0 or 1 and b is 1 or 2, provided that when a is 1 then b is 1;

c is 1 or 2, provided that when c is 1 then a is 0 and b is 1;

m is 1,2,3 or 4; and

p is 1 or 2;

or a pharmaceutically acceptable salt thereof.

As used herein, the term “alkyl” means a straight or branched C₁ to C₈carbon chain such as methyl, ethyl, tert-butyl, iso-propyl, n-octyl, andthe like. The term “lower alkyl” means a straight or branched C₁ to C₆carbon chain, such as methyl, ethyl, iso-propyl, and the like.

The term “cycloalkyl” means a mono-, bi-, or tricyclic ring that iseither fully saturated or partially unsaturated. Examples of such a ringinclude cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,adamantyl, cyclooctyl, cis- or trans decalin bicyclo[2.2.1]hept-2-ene,cyclohex-1-enyl, cyclopent-1-enyl, 1,4-cyclooctadienyl, and the like.

The term “(cycloalkyl)alkyl” means the above-defined alkyl groupsubstituted with one of the above cycloalkyl rings. Examples of such agroup include (cyclohexyl)methyl, 3-(cyclopropyl)-n-propyl,5-(cyclopentyl)hexyl, 6-(adamantyl)hexyl, and the like.

The term “substituted phenyl” specifies a phenyl group substituted withone or more substituents chosen from halogen, hydroxy, protectedhydroxy, cyano, nitro, trifluoromethyl, alkyl, alkoxy, acyl, acyloxy,carboxy, protected carboxy, carboxymethyl, protected carboxymethyl,hydroxymethyl, protected hydroxymethyl, amino, protected amino,(monosubstituted)amino, protected (monosubstituted)amino,(disubstituted)amino, carboxamide, protected carboxamide, N-(loweralkyl)carboxamide, protected N-(lower alkyl)carboxamide, N,N-di(loweralkyl)carboxamide, N-((lower alkyl)sulfonyl)amino,N-(phenylsulfonyl)amino or by a substituted or unsubstituted phenylgroup, such that in the latter case a biphenyl or naphthyl groupresults, or wherein two adjacent alkyl substituents on the phenyl ringtaken together from a cycloalkyl to yield, for example,teterahydronaphthyl or indanyl.

Examples of the term “substituted phenyl” includes a mono-, di-, tri-,tetra- or penta(halo)phenyl group such as 2-, 3- or 4-chlorophenyl,2,6-dichlorophenyl, 2,5-dichlorophenyl, 3,4-dichlorophenyl, 2-,3- or4-bromophenyl, 3,4-dibromophenyl, 3-chloro-4-fluorophenyl, 2-, 3- or4-fluorophenyl, 2,4,6-trifluorphenyl, 2,3,5,6-tetrafluorphenyl,2,3,4,5-tetrafluorophenyl, 2,3,4,5,6-pentafluoropheny, and the like; amono or di(hydroxy)phenyl group such as 2-, 3-, or 4-hydroxyphenyl,2,4-dihydroxyphenyl, the protected-hydroxy derivatives thereof and thelike; a nitrophenyl group such as 2-, 3-, or 4-nitrophenyl; acyanophenyl group, for example, 2-,3- or 4-cyanophenyl; a mono- ordi(alkyl)phenyl group such as 2-, 3-, or 4-methylphenyl,2,4-dimethylphenyl, 2-, 3- or 4-(iso-propyl)phenyl, 2-, 3-, or4-ethylphenyl, 2-, 3- or 4-(n-propyl)phenyl and the like; a mono ordi(alkoxy)phenyl group, for example, 2,6-dimethoxyphenyl, 2-, 3- or4-(iso-propoxy)phenyl, 2-, 3- or 4-(t-butoxy)phenyl,3-ethoxy-4-methoxyphenyl and the like; 2-, 3- or4-trifluoromethylphenyl; a mono- or dicarboxyphenyl or (protectedcarboxy)phenyl group such as 2-, 3- or 4-carboxyphenyl or2,4-di(protected carboxy)phenyl; a mono- or di(hydroxymethyl)phenyl or(protected hydroxymethyl)phenyl such as 2-, 3- or 4-(protectedhydroxymethyl)phenyl or 3,4-di(hydroxymethyl)phenyl; a mono- ordi(aminomethyl)phenyl or (protected aminomethyl)phenyl such as 2-, 3- or4-(aminomethyl)phenyl or 2,4-(protected aminomethyl)phenyl; or a mono-or di(N-(methylsulfonylamino))phenyl such as 2, 3 or4-(N-(methylsulfonylamino))phenyl. Also, the term “substituted phenyl”represents disubstituted phenyl groups wherein the substituents aredifferent, for example, 3-methyl-4-hydroxyphenyl,3-chloro-4-hydroxyphenyl, 2-methoxy-4-bromophenyl,4-ethyl-2-hydroxyphenyl, 3-hydroxy-4-nitrophenyl,2-hydroxy-4-chlorophenyl, and the like.

The term “phenylalkyl” means one of the above phenyl groups attached toone of the above-described alkyl groups, and the term “substitutedphenylalkyl means that either the phenyl or the alkyl, or both, aresubstituted with one or more of the above-identified substituents.Examples of such groups include 2-phenyl-1-chloroethyl,2-(4′-methoxyphenyl)ethyl, 4-(2′, 6′-dihydroxy phenyl)n-hexyl,2-(5′-cyano-3′-methoxyphenyl)n-pentyl, 3-(2′,6′-dimethylphenyl)n-propyl, 4-chloro-3-aminobenzyl,6-(4′-methoxyphenyl)-3-carboxy(n-hexyl),5-(4′-aminomethylphenyl)-3-(aminomethyl)n-pentyl,5-phenyl-3-oxo-n-pent-1-yl, (4-hydroxynapth-2-yl)methyl, and the like.

The term “substituted naphthyl” means a naphthyl group substituted withone or more of the above-identified substituents, and the term “(1 or 2naphyl)alkyl” means a naphthyl attached to one of the above-describedalkyl groups at the 1 or 2 position.

The terms “halo” and “halogen” refer to the fluoro, chloro, bromo oriodo groups. These terms may also be used to describe one or morehalogens, which are the same or different. Preferred halogens in thecontext of this invention are chloro and fluoro.

The term “aryl” refers to aromatic five and six membered carbocyclicrings. Six membered rings are preferred.

The term “heteroaryl” denotes optionally substituted aromaticfive-membered or six-membered heterocyclic rings that have 1 to 4heteroatoms, such as oxygen, sulfur and/or nitrogen atoms, in particularnitrogen, either alone or in conjunction with sulfur or oxygen ringatoms.

The following ring systems are representative examples of theheterocyclic radicals denoted by the term “heteroaryl” (whethersubstituted or unsubstituted): thienyl, furyl, pyrrolyl, pyrrolidinyl,imidazolyl, isoxazolyl, triazolyl, thiadiazolyl, oxadiazolyl,tetrazolyl, thiatriazolyl, oxatriazolyl, pyridyl, pyrimidyl, pyrazinyl,pyridazinyl, oxazinyl, triazinyl, thiadiazinyl tetrazolo,1,5-[b]pyridazinyl and purinyl, as well as benzo-fused derivatives, forexample, benzoxazolyl, benzothiazolyl, benzimidazolyl and indolyl.

Substituents for the above optionally substituted heteroaryl rings arefrom one to three halo, trihalomethyl, amino, protected amino, aminosalts, mono-substituted amino, di-substituted amino, carboxy, protectedcarboxy, carboxylate salts, hydroxy, protected hydroxy, salts of ahydroxy group, lower alkoxy, lower alkylthio, lower alkyl, substitutedalkyl, cycloalkyl, substituted cycloalkyl, (cycloalkyl)alkyl,substituted (cycloalkyl)alkyl, phenyl, substituted phenyl, phenylalkyl,and substituted phenylalkyl groups.

Substituents for the heteroaryl group are as defined above, or as setforth below. As used in conjunction with the above substituents forheteroaryl rings, “trihalomethyl” can be trifluoromethyl,trichloromethyl, tribromomethyl or triiodomethyl, “lower alkoxy” means aC₁ to C₄ alkoxy group, similarly, “lower alkylthio” means a C₁ to C₄alkylthio group. The term “substituted lower alkyl” means theabove-defined lower alkyl group substituted from one to three times by ahydroxy, protected hydroxy, amino, protected amino, cyano, halo,trifluoromethyl, mono-substituted amino, di-substituted amino, loweralkoxy, lower alkylthio, carboxy, protected carboxy, or a carboxy,amino, and/or hydroxy salt.

As used in conjunction with the substituents for the heteroaryl rings,the terms “substituted (cycloalkyl)alkyl” and “substituted cycloalkyl”are as defined above substituted with the same groups as listed for a“substituted alkyl” group. The term “(monosubstituted)amino” refers toan amino group with one substituent chosen from the group consisting ofphenyl, substituted phenyl, alkyl, substituted alkyl, C₁ to C₇ acyl, C₂to C₇ alkenyl, C₂ to C₇ substituted alkenyl, C₂ to C₇ alkynyl, C₇ to C₁₆alkylaryl, C₇ to C₁₆ substituted alkylaryl and heteroaryl group. The(monosubstituted)amino can additionally have an amino-protecting groupas encompassed by the term “protected (monosubstituted)amino.” The term“(disubstituted)amino” refers to amino groups with two substituentschosen from the group consisting of phenyl, substituted phenyl, alkyl,substituted alkyl, C₁ to C₇ acyl, C₂ to C₇ alkenyl, C₂ to C₇ alkynyl, C₇to C₁₆ alkylaryl, C₇ to C₁₆ substituted alkylaryl and heteroaryl. Thetwo substituents can be the same or different. The term“heteroaryl(alkyl)” denotes an alkyl group as defined above, substitutedat any position by a heteroaryl group, as above defined.

Furthermore, the above optionally substituted five-membered orsix-membered heterocyclic rings can optionally be fused to a aromatic5-membered or 6-membered aryl or heteroaryl ring system. For example,the rings can be optionally fused to an aromatic 5-membered or6-membered ring system such as a pyridine or a triazole system, andpreferably to a benzene ring.

The term “pharmaceutically-acceptable salt” encompasses those salts thatform with the carboxylate anions and includes salts formed with theorganic and inorganic cations such as those chosen from the alkali andalkaline earth metals, (for example, lithium, sodium, potassium,magnesium, barium and calcium); and ammonium ion; and the organiccations (for example, dibenzylammonium, benzylammonium,2-hydroxyethylammonium, bis(2-hydroxyethyl)ammonium,phenylethylbenzylammonium, dibenzylethylenediammonium, and likecations.) Other cations encompassed by the above term include theprotonated form of procaine, quinine and N-methylglucosamine, theprotonated forms of basic amino acids such as glycine, ornithine,histidine, phenylglycine, lysine, and arginine. Furthermore, anyzwitterionic form of the instant compounds formed by a carboxylic acidand an amino group is referred to by this term. A preferred cation forthe carboxylate anion is the sodium cation. Furthermore, the termincludes salts that form by standard acid-base reactions with basicgroups (such as amino groups) and includes organic or inorganic acids.Such acids include hydrochloric, sulfuric, phosphoric, acetic, succinic,citric, lactic, maleic, fumaric, palmitic, cholic, pamoic, mucic,D-glutamic, D-camphoric, glutaric, phthalic, tartaric, lauric, stearic,salicyclic, methanesulfonic, benzenesulfonic, sorbic, picric, benzoic,cinnamic, and the like acids.

The compounds of Formula I may also exist as solvates and hydrates.Thus, these compounds may crystallize with, for example, waters ofhydration, or one, a number of, or any fraction thereof of molecules ofthe mother liquor solvent. The solvates and hydrates of such compoundsare included within the scope of this invention.

The term “carboxy-protecting group” as used herein refers to one of theester derivatives of the carboxylic acid group commonly employed toblock or protect the carboxylic acid group while reactions are carriedout on other functional groups on the compound. Examples of suchcarboxylic acid protecting groups include t-butyl, 4-nitrobenzyl,4-methoxybenzyl, 3,4-dimethoxybenzyl, 2,4-dimethoxybenzyl,2,4,6-trimethoxybenzyl, 2,4,6-trimethylbenzyl, pentamethylbenzyl,3,4-methylenedioxybenzyl, benzhydryl, 4,4′-dimethoxytrityl, 4,4′,4″-trimethoxytrityl, 2-phenylpropyl, trimethylsilyl,t-butyldimethylsilyl, phenacyl, 2,2,2-trichloroethyl,β-(trimethylsilyl)ethyl, β-(di(n-butyl)methylsilyl)ethyl,p-toluenesulfonylethyl, 4-nitrobenzylsulfonylethyl, allyl, cinnamyl,1-(trimethylsilylmethyl)-propenyl and like moieties. The species ofcarboxy-protecting group employed is not critical so long as thederivatized carboxylic acid is stable to the conditions of subsequentreaction(s) and can be removed at the appropriate point withoutdisrupting the remainder of the molecule. Further examples of thesegroups are found in C. B. Reese and E. Haslam, “Protective Groups inOrganic Chemistry,” J. G. W. McOmie, Ed., Plenum Press, New York, N.Y.,1973, Chapter 5, respectively, and T. W. Greene and P. G. M. Wuts,“Protective Groups in Organic Synthesis,” 2nd ed., John Wiley and Sons,New York, N.Y., 1991, Chapter 5, each of which is incorporated herein byreference. A related term is “protected carboxy,” which refers to acarboxy group substituted with one of the above carboxy-protectinggroups.

The term “hydroxy-protecting group” refers to readily cleavable groupsbonded to hydroxyl groups, such as the tetrahydropyranyl,2-methoxyprop-2-yl, 1-ethoxyeth-1-yl, methoxymethyl,β-methoxyethoxymethyl, methylthiomethyl, t-butyl, t-amyl, trityl,4-methoxytrityl, 4,4′-dimethoxytrityl, 4,4′,4″-trimethoxytrityl, benzyl,allyl, trimethylsilyl, (t-butyl)dimethylsilyl,2,2,2-trichloroethoxycarbonyl, and the like.

Further examples of hydroxy-protecting groups are described by C. B.Reese and E. Haslam, “Protective Groups in Organic Chemistry,” J. G. W.McOmie, Ed., Plenum Press, New York, N.Y., 1973, Chapters 3 and 4,respectively, and T. W. Greene and P. G. M. Wuts, “Protective Groups inOrganic Synthesis,” Second Edition, John Wiley and Sons, New York, N.Y.,1991, Chapters 2 and 3. A preferred hydroxy-protecting group is thetert-butyl group. The related term “protected hydroxy” denotes a hydroxygroup bonded to one of the above hydroxy-protecting groups.

The term “amino-protecting group” as used herein refers to substituentsof the amino group commonly employed to block or protect the aminofunctionality while reacting other functional groups of the molecule.The term “protected (monosubstituted)amino” means there is anamino-protecting group on the monosubstituted amino nitrogen atom.

Examples of such amino-protecting groups include the formyl (“For”)group, the trityl group, the phthalimido group, the trichloroacetylgroup, the trifluoroacetyl group, the chloroacetyl, bromoacetyl, andiodoacetyl groups, urethane-type protecting groups, such ast-butoxycarbonyl (“Boc”), 2-(4-biphenylyl)propyl-2-oxycarbonyl (“Bpoc”),2-phenylpropyl-2-oxycarbonyl (“Poc”), 2-(4-xenyl)isopropoxycarbonyl,1,1-diphenylethyl-1-oxycarbonyl, 1,1-diphenylpropyl-1-oxycarbonyl,2-(3,5-dimethoxyphenyl)propyl-2-oxycarbonyl (“Ddz”),2-(p-toluyl)propyl-2-oxycarbonyl, cyclopentanyloxycarbonyl,1-methylcyclopentanyl-oxycarbonyl, cyclohexanyloxy-carbonyl,1-methyl-cyclohexanyloxycarbonyl, 2-methylcyclohexanyl-oxycarbonyl,2-(4-toluylsulfonyl)ethoxycarbonyl, 2-(methylsulfonyl)ethoxycarbonyl,2-(triphenylphosphino)-ethoxycarbonyl, 9-fluorenylmethoxycarbonyl(“Fmoc”), 2-(trimethylsilyl)ethoxycarbonyl, allyloxycarbonyl,1-(trimethylsilylmethyl)prop-1-enyloxycarbonyl,5-benzisoxalylmethoxycarbonyl, 4-acetoxybenzyl-oxycarbonyl,2,2,2-trichloroethoxycarbonyl, 2-ethynyl-2-propoxycarbonyl,cyclopropylmethoxycarbonyl, isobornyloxycarbonyl,1-piperidyloxycarbonyl, benzyloxycarbonyl (“Cbz”),4-phenylbenzyloxycarbonyl, 2-methylbenzyloxycarbonyl,α-2,4,5,-tetramethylbenzyl-oxycarbonyl (“Tmz”),4-methoxybenzyloxycarbonyl, 4-fluorobenzyloxycarbonyl,4-chlorobenzyloxycarbonyl, 3-chlorobenzyloxycarbonyl,2-chlorobenzyloxycarbonyl, 2,4-dichlorobenzyloxycarbonyl,4-bromobenzyloxycarbonyl, 3-bromobenzyloxycarbonyl,4-nitrobenzyloxycarbonyl, 4-cyanobenzyloxycarbonyl,4-(decyloxy)benzyloxycarbonyl and the like; the benzoylmethylsulfonylgroup, the 2,2,5,7,8-pentamethylchroman-6-sulfonyl group (“PMC”), thedithiasuccinoyl (“Dts”) group, the 2-(nitro)phenyl-sulfenyl group(“Nps”), the diphenylphosphine oxide group, and like amino-protectinggroups. The species of amino-protecting group employed is not criticalso long as the derivatized amino group is stable to the conditions ofthe subsequent reaction(s) and can be removed at the appropriate pointwithout disrupting the remainder of the molecule. Preferredamino-protecting groups are Boc, Cbz and Fmoc. Further examples ofamino-protecting groups embraced by the above term are well known inorganic synthesis and the peptide art and are described by, for example,T. W. Greene and P. G. M. Wuts, “Protective Groups in OrganicSynthesis,” 2nd ed., John Wiley and Sons, New York, N.Y., 1991, Chapter7, M. Bodanzsky, “Principles of Peptide Synthesis,” 1st and 2nd revisedEd., Springer-Verlag, New York, N.Y., 1984 and 1993, and J. M. Stewartand J. D. Young, “Solid Phase Peptide Synthesis,” 2nd Ed., PierceChemical Co., Rockford, Ill., 1984, E. Atherton and R. C. Shephard,“Solid Phase Peptide Synthesis—A Practical Approach” IRL Press, Oxford,England (1989), each of which is incorporated herein by reference. Therelated term “protected amino” defines an amino group substituted withan amino-protecting group discussed above.

The terms “natural and unnatural amino acid” refers to both thenaturally occurring amino acids and other non-proteinogenic α-aminoacids commonly utilized by those in the peptide chemistry arts whenpreparing synthetic analogues of naturally occurring peptides, includingD and L forms. The naturally occurring amino acids are glycine, alanine,valine, leucine, isoleucine, serine, methionine, threonine,phenylalanine, tyrosine, tryptophan, cysteine, proline, histidine,aspartic acid, asparagine, glutamic acid, glutamine, γ-carboxyglutamicacid, arginine, ornithine and lysine. Examples of unnatural alpha-aminoacids include hydroxylysine, citrulline, kynurenine,(4-aminophenyl)alanine, 3-(2′-naphthyl)alanine, 3-(1′-naphthyl)alanine,methionine sulfone, (t-butyl)alanine, (t-butyl)glycine,4-hydroxyphenyl-glycine, aminoalanine, phenylglycine, vinylalanine,propargyl-gylcine, 1,2,4-triazolo-3-alanine, thyronine,6-hydroxytryptophan, 5-hydroxytryptophan, 3-hydroxy-kynurenine,3-aminotyrosine, trifluoromethylalanine, 2-thienylalanine,(2-(4-pyridyl)ethyl)cysteine, 3,4-dimethoxy-phenylalanine,3-(2′-thiazolyl)alanine, ibotenic acid,1-amino-1-cyclopentane-carboxylic acid, 1-amino-1-cyclohexanecarboxylicacid, quisqualic acid, 3-(trifluoromethylphenyl)alanine,(cyclohexyl)glycine, thiohistidine, 3-methoxytyrosine, norleucine,norvaline, alloisoleucine, homoarginine, thioproline, dehydro-proline,hydroxyproline, homoproline, indoline-2-carboxylic acid,1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid,1,2,3,4-tetrahydroquinoline-2-carboxylic acid, α-amino-n-butyric acid,cyclohexylalanine, 2-amino-3-phenylbutyric acid, phenylalaninesubstituted at the ortho, meta, or para position of the phenyl moietywith one or two of the following groups: a (C₁ to C₄)alkyl, a (C₁ toC₄)alkoxy, a halogen or a nitro group, or substituted once with amethylenedioxy group; β-2- and 3-thienylalanine; β-2- and3-furanylalanine; β-2-, 3- and 4-pyridylalanine; β-(benzothienyl-2- and3-yl)alanine; β-(1- and 2-naphthyl)alanine; O-alkylated derivatives ofserine, threonine or tyrosine; S-alkylated cysteine, S-alkylatedhomocysteine, the O-sulfate, O-phosphate and O-carboxylate esters oftyrosine; 3-(sulfo)tyrosine, 3-(carboxy)tyrosine, 3-(phospho)tyrosine,the 4-methane-sulfonic acid ester of tyrosine, 4-methanephosphonic acidester of tyrosine, 3,5-diiodotyrosine, 3-nitrotyrosine, ε-alkyllysine,and delta-alkyl ornithine. Any of these α-amino acids may be substitutedwith a methyl group at the alpha position, a halogen at any position ofthe aromatic residue on the α-amino side chain, or an appropriateprotective group at the O, N, or S atoms of the side chain residues.Appropriate protective groups are discussed above.

The compounds of this invention may be modified by appropriatefunctionalities to enhance selective biological properties. Suchmodifications are known in the art and include those which increasebiological penetration into a given biological system (e.g., blood,lymphatic system, central nervous system), increase oral availability,increase solubility to allow administration by injection, altermetabolism and alter rate of exertion. In addition, the compounds may bealtered to pro-drug form such that the desired compound is created inthe body of the patient as the result of the action of metabolic orother biochemical processes on the pro-drug. Some examples of pro-drugforms include ketal, acetal, oxime, and hydrazone forms of compoundswhich contain ketone or aldehyde groups, especially where they occur inthe group donated as “A” in Formula I or the modified aspartic acidresidue attached to the group denoted as “A”.

With regard to the q and r groups of Formula I, typical embodimentsinclude compounds wherein q is 1 and r is 2.

Compounds of this invention with respect to the n, R, R¹, R² and Xgroups in Formula I include those wherein:

R is lower alkyl (such as methyl);

R¹ is substituted phenyl (such as 2-substituted phenyl), naphthyl, orsubstituted naphthyl;

R² is hydrogen, lower alkyl, (CH₂)_(p)CO₂R³, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or 2-naphthyl), or(CH₂)_(m)tetrazolyl,

where p is 1 or 2, m is 1 or 2;

X is O or NH; and

n is 0 or 1.

Other compounds of this invention with respect to the R¹, R² and Xgroups in Formula I include those wherein:

R¹ is substituted phenyl, naphthyl, or substituted naphthyl;

R₂ is (CH₂)_(m)tetrazolyl, where m is 1 or 2; and

X is C═ONH.

Compounds of this invention with respect to the A group in Formula Iinclude those of Formula IIa wherein:

R⁴ is lower alkyl, cycloalkyl, phenyl, substituted phenyl, (CH₂)_(m)NH₂,(CH₂)_(m)OR¹¹, (CH₂)_(p)SR¹², (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), or (CH₂)_(m)(1 or 2-naphthyl);

R¹¹ is hydrogen, lower alkyl, cycloalkyl, phenyl, substituted phenyl,naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substitutedphenyl), or (CH₂)_(m)(1 or 2-naphthyl);

R¹² is lower alkyl, cycloalkyl, phenyl, substituted phenyl, naphthyl,(CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), or(CH₂)_(m)(1 or 2-naphthyl); and

m is 1,2,3,4 and p is 1 or2.

Compounds of this invention with respect to the A group in Formula Ialso include those of Formula IIb wherein:

R⁵ is phenyl, substituted phenyl, (CH₂)_(p)phenyl, (CH₂)_(p)(substitutedphenyl), cycloalkyl, or 2-indanyl; and

p is 1 or 2.

Another group of compounds with respect to the A group in Formula I,include those of Formula IId wherein:

R⁷ is hydrogen, fluorine, cycloalkyl, phenyl, substituted phenyl,naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substitutedphenyl), (CH₂)_(m)(1 or 2-naphthyl), OR¹¹, or SR¹²;

R¹¹ and R¹² are independently cycloalkyl, phenyl, substituted phenyl,naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substitutedphenyl), or (CH₂)_(m)(1 or 2-naphthyl); and

m is 1,2,3 or 4.

A forth group of compounds with respect to the A group in Formula Iinclude those of Formula IIe wherein:

R⁸ is hydrogen, oxo, cycloalkyl, phenyl, substituted phenyl, ornaphthyl; and

Y¹ is CH₂, (CH₂)₂, (CH₂)₃, or S.

Another group of compounds with respect to the A group in Formula Iinclude those of Formula IIh wherein:

a is 0 and b is 1 or 2.

Compounds of this invention with respect to the B group in Formula Iinclude those wherein:

B is hydrogen, 2-benzoxazolyl, substituted 2-oxazolyl, CH₂ZR¹⁶,CH₂OCO(aryl), or CH₂OPO(R¹⁷)R¹⁸, where Z is O or S;

R¹⁶is phenyl, substituted phenyl, naphthyl, substituted naphthyl,heteroaryl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1or 2-naphthyl), or (CH₂)_(m)heteroaryl; and

R¹⁷ and R¹⁸ are independently alkyl, cycloalkyl, phenyl, substitutedphenyl, naphthyl, phenylalkyl, substituted phenylalkyl and(cycloalkyl)alkyl.

Another group of compounds with respect to the B group in Formula Iinclude those of Formula IIIa-c wherein:

Y² is O or NR²⁴;

Y³ is CH₂, O, or NR²⁴;

R¹⁹ and R²⁰ are independently hydrogen, alkyl, phenyl, or R¹⁹ and R²⁰taken together are —(CH═CH)₂—;

R²¹ is hydrogen, alkyl, phenyl, substituted phenyl, (CH₂)_(m)phenyl, or(CH₂)_(m)(substituted phenyl); and

R²², R²³ and R²⁴ are independently hydrogen or alkyl.

The compounds of Formula I may be synthesized using conventionaltechniques as discussed below. Advantageously, these compounds areconveniently synthesized from readily available starting materials toform a carboxylic acid intermediate as represented in the followingReaction Schemes 1 and 2. To this end, in the following syntheticschemes, q is 1, and corresponding compounds wherein q is 2 may be madein the same manner by employing the corresponding ethylene (—CH₂CH₂—)starting material in place of the methylene (—CH₂—) moiety.

One synthetic route for synthesizing the carboxylic acid intermediate isset forth in the following Scheme 1:

In the above Scheme 1, R′ represents hydrogen or a carboxy-protectinggroup, wherein the carboxy-protecting group is as defined above. “PG”stands for an amino-protecting group, and “A” stands for a natural orunnatural amino acid of Formula IIa through IIi, as discussed above. Inaddition, Formula (V) above (i.e., H₂N—C) represents a modified asparticacid residue of Formulas Va through Vd:

The modified aspartic acids of Formula Va-d can be prepared by methodswell known in the art. See, for example, European Patent Application519,748; PCT Patent Application No. PCT/EP92/02472; PCT PatentApplication No. PCT/US91/06595; PCT Patent Application No.PCT/US91/02339; European Patent Application No. 623,592; World PatentApplication No. WO 93/09135; PCT Patent Application No. PCT/US94/08868;European Patent Application No. 623,606; European Patent Application No.618,223; European Patent Application No. 533,226; European PatentApplication No. 528,487; European Patent Application No. 618,233; PCTPatent Application No. PCT/EP92/02472; World Patent Application No. WO93/09135; PCT Patent Application No. PCT/US93/03589; and PCT PatentApplication No. PCT/US93/00481, all of which are herein incorporated byreference.

The coupling reactions carried out under Step A are performed in thepresence of a standard peptide coupling agent such as the combination ofthe combination of dicyclohexylcarbodiimide(DCC) and1-hydroxy-benzotriazole(HOBt), as well as the BOP(benzotriazolyloxy-tris-(dimethylamino)phosphonium hexafluorophosphate)reagent, pyBOP(benzotriazolyloxy-tris(N-pyrolidinyl)phosphoniumhexafluorophosphate),HBTU (O-benzotriazolyly-tetramethylisouronium-hexafluorophosphate), andEEDQ (1-ethyloxycarbonyl-2-ethyloxy-1,2-dihydroquinoline) reagents, thecombination of 1-ethyl(3,3′-dimethyl-1′-aminopropyl)carbodiimide (EDAC)and HOBt, and the like, as discussed in J. Jones, “Amino Acid andPeptide Synthesis,” Steven G. Davis ed., Oxford University Press,Oxford, pp. 25-41 (1992); M. Bodanzky, “Principles of PeptideSynthesis,” Hafher et al. ed., Springer-Verlag, Berlin Heidelberg, pp.9-52 and pp. 202-251 (1984); M. Bodanzky, “Peptide Chemistry, APractical Textbook,” Springer-Verlag, Berlin Heidelberg, pp. 55-73 andpp. 129-180; and Stewart and Young, “Solid Phase Peptide Synthesis,”Pierce Chemical Company, (1984), all of which are herein incorporated byreference. The amino protecting group is then removed and the resultingamine is coupled to the (substituted) carboxylic acid of Formula VII(Step B). Again, this coupling reaction uses the standard peptidecoupling reactions mentioned above.

Alternatively, the (substituted)carboxylic acid of Formula VII can becoupled to an amino ester of Formula IX (Step D). Again, this couplingreaction uses the standard peptide coupling reactions mentioned above.In Formula IX, the group R is a carboxyl protecting group such asmethyl, allyl, benzyl or tert-butyl. After removal of the carboxylprotecting group under standard conditions well known in the art, theresulting carboxylic acid is coupled to amine V using the standardpeptide coupling methods described above (Step E).

In the case where the coupling reaction depicted by either Step A orStep E was carried out with the amino alcohol of Formula Vc, the alcoholmoiety must be oxidized to the corresponding carbonyl compound prior toremoval of the protecting groups. Preferred methods for the oxidationreaction include Swern oxidation (oxalyl chloride-dimethyl sulfoxide,methylene chloride at −78° C. followed by triethylamine); andDess-Martin oxidation (Dess-Martin periodinane, t-butanol, and methylenechloride.) The protecting groups contained in substructures of theFormula Va-d, VII and A are removed by methods well known in the art.These reactions and removal of some or all of the protecting groups areinvolved in Step C in the above Scheme 1.

An alternative synthetic route for synthesizing the carboxylig acidintermediate having a protected carboxy group is set forth in thefollowing Scheme 2:

In the above Scheme 2, “PG” stands for an amino protecting group and “A”stands for a natural or unnatural amino acid of formula IIa through IIi,as discussed above. The R′ is a carboxyl protecting group such astrimethylsilyl, methyl, allyl, benzyl or tert-butyl.

The coupling reactions carried out under Step F and Step G are performedin the presence of a standard peptide coupling agent as discussed above.In Step G, the amino protecting group must be removed prior to thecoupling step. In Step H the alpha-carboxy protecting group R of thecompound of Formula XIII is selectively removed and the resultingmono-carboxylic acid treated sequentially with diazomethane andhydrobromic acid to give the alpha-bromoketone of Formula XIV.

In Step I, the bromoketone of Formula XIV is treated with either R¹⁶Z—H,(aryl)-CO₂H, (heteroaryl)-CO₂H, or R¹⁷(R¹⁸)PO₂H in the presence of aninorganic base such as potassium carbonate or potassium fluoride in aninert solvent such as dimethyl formamide to give the correspondingcompound of Formula Ia in which B is CH₂ZR¹⁶, CH₂OCO(aryl),CH₂OCO(heteroaryl), or CH₂OPO(R¹⁷)R¹⁸, respectively. Compounds ofFormula Ia in which B is a fragment of Formula III may also be preparedin a similar fashion. The protecting groups contained in substructuresof the Formula VII, XI and A are removed by methods well known in theart. These reactions and removal of some or all of the protecting groupsare involved in Step I in the above Scheme 2.

An alternative method for the preparation of the carboxylic acidintermediate of Formula Ia in which R′ and B are both hydrogen (i.e.,Formula Ib below) is set forth in Scheme 3:

In Scheme 3, Fmoc is the amino protecting group9-fluorenylmethoxycarbonyl and the shaded circle labeled “PS” representspolystyrene resin.

The coupling of the acid of Formula XV to a primary amine on solidsupport, preferably aminomethyl polystyrene, is carried out usingstandard peptide coupling agents, preferably usingbenzotriazolyloxy-tris(N-pyrolidinyl)phosphoniumhexafluorophosphate(pyBOP) in a inert solvent such as dimethylformamide or N-methylpyrrolidone (Step J). After removal of the Fmoc protecting group of XVIby treatment with pyrrolidine-dimethylformamide, the resulting amine iscoupled to Fmoc-amino acid of Formula IVa using standard peptidecoupling conditions as discussed above (Step K).

In Step L the Fmoc protecting group of the compound of Formula XVII isremoved again by treatment with pyrrolidine-dimethylformamide and theresulting amine coupled to the (substituted)carboxylic acid of FormulaVII again using standard peptide coupling conditions as discussed above.The tert-butyl ester of the compound of Formula XVIII is removed bytreatment with trfluoroacetic acid-methylene chloride in the presence ofa trapping agent such as anisole and the resulting acid cleaved from thesolid support by treatment with 37% aqueous formaldehyde/aceticacid/tetrahydrofuran/trifluoroacetic acid, preferably in a ratio of1/1/5/0.025, to give the aspartyl aldehyde of Formula Ib (Step M).

Once synthesized by, for example, the above techniques, the carboxylicacid intermediate of Formula Ia (R′═H) may then be converted tocompounds of Formula I according to the following Reaction Scheme 4:

In Reaction Scheme 4, conversion of the carboxylic acid intermediate tothe corresponding sulfonimide is typically accomplished by utilizing anintermediate having a protected carboxyl group through which the Bmoiety is attached. For example, in place of the —C(═O)B moiety, ahydroxy-protected group may be employed, such as —C(OTHP)B. Asrepresented by Step N in Reaction Scheme 5, this hydroxy-protectedintermediate, Formula Ic, may be converted to the correspondingsulfonimide intermediate of Formula Id by treatment with CDI (2 eq.) inTHF at room temperature for 3 hours, followed by H₂NS(O)_(r)R (2 eq.) inDBU (2 eq.) at room temperature for 4 hours.

The sulfonimide intermediate of Formula Id is reacted in Step O withTsOH (0.4 eq.) in methanol at room temperature for 30 minutes tode-protect the alcohol of Formula Ie, which in Step P may be convertedto the corresponding carbonyl of Formula I by employing the Dess-Martinperiodinane reagent and DCM at room temperature for 30 minutes.

Alternatively, a stabilize sulfonamide ring may first be formed and thenadded to the remainder of the molecule via amide bond formation with thecarboxy terminus of the natural or unnatural amino acid A, asillustrated in Reaction Scheme 6

This reaction is further illustrated in Example 3 below.

Depending on the choice of solvent and other conditions known to thepractitioner skilled in the art, compounds of this invention may alsotake a cyclized form, which forms are included in the instant invention.In particular, when B is hydrogen compounds of Formula I may exist inthe cyclic Formula I′ shown below:

When B is a moiety other than hydrogen, and depending upon the choice ofsolvents (e.g., ROH), the compounds of the cyclic form also includecompounds having Formula I″ as shown below.

In addition, it should be understood that the equilibrium forms of thecompounds of this invention may include tautomeric forms. All such formsof these compounds are expressly included in the present invention.

Pharmaceutical compositions of this invention comprise any of thecompounds of the present invention, and pharmaceutically acceptablesalts thereof, with any pharmaceutically acceptable carrier, adjuvant orvehicle (hereinafter collectively referred to as“pharmaceutically-acceptable carriers”). Pharmaceutically acceptablecarriers, adjuvants and vehicles that may be used in the pharmaceuticalcompositions of this invention include, but are not limited to, ionexchange, alumina, aluminum stearate, lecithin, serum proteins, such ashuman serum albumin; buffer substances such as the various phosphates,glycine, sorbic acid, potassium sorbate, partial glyceride mixtures ofsaturated vegetable fatty acids; water, salts or electrolytes, such asprotamine sulfate, disodium hydrogen phosphate, potassium hydrogenphosphate, sodium chloride, and zinc salts; colloidal silica, magnesiumtrisilicate, polyvinyl pyrrolidone, cellulose-based substances,polyethylene glycol, sodium carboxymethylcellulose, polyarylates, waxes,polyethylene-polyoxypropylene-block polymers, polyethylene glycol andwool fat, and the like.

The pharmaceutical compositions of this invention may be administeredorally, parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or by an implanted reservoir. Oral and parenteraladministration are preferred. The term “parenteral” as used hereinincludes subcutaneous, intracutaneous, intravenous, intramuscular,intra-articular, intrasynovial, intrasternal, intrathecal, intralesionaland intracranial injection or infusion techniques.

The pharmaceutical compositions may be in the form of a sterileinjectable preparation, for example, as a sterile injectable aqueous oroleaginous suspension. This suspension may be formulated according totechniques known in the art using suitable dispersing or wetting agents(such as, for example, Tween 80) and suspending agents. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally-acceptable diluent or solvent,for example, as a solution in 1,3-butanediol. Among the acceptablevehicles and solvents that may be employed are mannitol, water, Ringer'ssolution and isotonic sodium chloride solution. In addition, sterile,fixed oils are conventionally employed as a solvent or suspendingmedium. For this purpose, any bland fixed oil may be employed includingsynthetic mono- or diglycerides. Fatty acids, such as oleic acid and itsglyceride derivatives are useful in the preparation of injectables, asare natural pharmaceutically-acceptable oils, such as olive oil orcastor oil, especially in their polyoxyethylated versions. These oilsolutions or suspensions may also contain a long-chain alcohol diluentor dispersant.

The pharmaceutical compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, and aqueous suspensions and solutions. Inthe case of tablets for oral use, carrier which are commonly usedinclude lactose and corn starch. Lubricating agents, such as magnesiumstearate, are also typically added. For oral administration in capsuleform useful diluents include lactose and dried corn starch. When aqueoussuspensions are administered orally, the active ingredient is combinedwith emulsifying and suspending agents. If desired, certain sweeteningand/or flavoring and/or coloring agents may be added.

The pharmaceutical compositions of this invention may also beadministered in the form of suppositories for rectal administration.These compositions can be prepared by mixing a compound of thisinvention with a suitable non-irritating excipient which is solid atroom temperature but liquid at the rectal temperature. Such materialsinclude, but are not limited to, cocoa butter, beeswax and polyethyleneglycols.

Topical administration of the pharmaceutical compositions of thisinvention is especially useful when the desired treatment involves areasor organs readily accessible to topical application. For applicationtopically to the skin, the pharmaceutical composition should beformulated with a suitable ointment containing the active componentssuspended or dissolved in a carrier. Carriers for topical administrationof the compounds of this invention include, but are not limited to,mineral oil, liquid petroleum, white petroleum, propylene glycol,polyoxyethylene, polyoxypropylene compound, emulsifying wax and water.Alternatively, the pharmaceutical composition can be formulated with asuitable lotion or cream containing the active compound suspended ordissolved in a carrier. Suitable carriers include, but are not limitedto, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esterswax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water. Thepharmaceutical compositions of this invention may also be topicallyapplied to the lower intestinal tract by rectal suppository formulationor in a suitable enema formulation. Topically-applied transdermalpatches are also included in this invention.

The pharmaceutical compositions of this invention may be administered bynasal aerosol or inhalation. Such compositions are prepared according totechniques well-known in the art of pharmaceutical formulation and maybe prepared as solutions in saline, employing benzyl alcohol or othersuitable preservatives, absorption promoters to enhance bioavailability,fluorocarbons, and/or other solubilizing or dispersing agents known inthe art.

The compounds of this invention may be used in combination with eitherconventional anti-inflammatory agents or with matrix metalloproteaseinhibitors, lipoxygenase inhibitors and antagonists of cytokines otherthan IL-1β.

The compounds of this invention can also be administered in combinationwith immunomodulators (erg., bropirimine, anti-human alpha interferonantibody, IL-2, GM-CSF, methionine enkephalin, interferon alpha,diethyldithiocarbamate, tumor necrosis factor, naltrexons and rEPO) orwith prostaglandins, to prevent or combat IL-1-mediated disease symptomssuch as inflammation.

When the compounds of this invention are administered in combinationtherapies with other agents, they may be administered sequentially orconcurrently to the patient. Alternatively, pharmaceutical compositionsaccording to this invention may be comprised of a combination of acompound of Formula I and another therapeutic or prophylactic agentmentioned above.

The disease states which may be treated or prevented by the instantpharmaceutical compositions include, but are not limited to,inflammatory diseases, autoimmune diseases and neurodegenerativediseases, and for inhibiting unwanted apoptosis involved in ischemicinjury, such as ischemic injury to the heart (e.g., myocardialinfarction), brain (e.g., stroke), and kidney (e.g., ischemic kidneydisease). As a consequence of their ability to inhibit apoptosis, thepresent pharmaceutical compositions are also useful for the repopulationof hematopoietic cells of a patient following chemotherapy. Methods ofadministering an effective amount of the above-described pharmaceuticalcompositions to mammals, also referred to herein as patients, in need ofsuch treatment (that is, those suffering from inflammatory diseases,autoimmune diseases, neurodegenerative diseases and for the repopulationof hematopoictic cells in cancer patients who have undergonechemotherapy) are another aspect of the instant invention. Finally, as afurther consequence of their ability to inhibit apoptosis, the instantpharmaceutical compositions may be used in a method to prolong theviability of organs to be used in transplantations.

Inflammatory disease which may be treated or prevented include, forexample, septic shock, septicemia, and adult respiratory distresssyndrome. Target autoimmune diseases include, for example, rheumatoid,arthritis, systemic lupus erythematosus, scleroderma, chronicthyroiditis, Graves' disease, autoimmune gastritis, insulin-dependentdiabetes mellitus, autoimmune hemolytic anemia, autoimmune neutropenia,thrombocytopenia, chronic active hepatitis, myasthenia gravis andmultiple sclerosis. Target neurodegenerative diseases include, forexample, amyotrophic lateral sclerosis, Alzheimer's disease, Parkinson'sdisease, and primary lateral sclerosis. The pharmaceutical compositionsof this invention may also be used to promote wound healing. Targetdiseases associated with harmful, apoptosis, in other words, thoseassociated with ischemic injury, includes myocardial infarction, stroke,and ischemic kidney disease. The pharmaceutical compositions of thisinvention may also be used to treat infectious diseases, especiallythose involved with viral infections.

The term “effective amount” refers to dosage levels of the order of fromabout 0.05 milligrams to about 140 milligrams per kilogram of bodyweight per day for use in the treatment of the above-indicatedconditions (typically about 2.5 milligrams to about 7 grams per patientper day). For example, inflammation may be effectively treated by theadministration of from about 0.01 to 50 milligrams of the compound perkilogram of body weight per day (about 0.5 milligrams to about 3.5 gramsper patient per day).

The amount of the compounds of Formula I that may be combined with thecarrier materials to produce a single dosage form will vary dependingupon the host treated and the particular mode of administration. Forexample, a formulation intended for the oral administration of humansmay contain from 0.5 milligrams to 5 grams of a compound of Formula Icombined with an appropriate and convenient amount of apharmaceutically-acceptable carrier which may vary from about 5 to about95 percent of the total composition. Dosage unit forms will generallycontain between from about 1 milligram to about 500 milligrams of anactive compound of Formula I.

It will be understood, however, that the specific “effective amount” forany particular patient will depend upon a variety of factors includingthe activity of the specific compound employed, the age, body weight,general health, sex, diet, time of administration, route ofadministration, rate of excretion, drug combination and the severity ofthe particular disease undergoing prevention or therapy.

Although this invention focuses on the use of the compounds disclosedherein for preventing and treating IL-1-mediated diseases, the compoundsof this invention can also be used as inhibitory agents for othercysteine proteases.

The compounds of this invention are also useful as commercial reagentswhich effectively bind to the ICE/ced-3 family of cysteine protease orother cysteine proteases. As commercial reagents, the compounds of thisinvention, and their derivatives, may be used to block proteolysis of atarget peptide or may be derivatized to bind to a stable resin as atethered substrate for affinity chromatography applications. These andother uses which characterize commercial cystine protease inhibitorswill be evident to those of ordinary skill in the art.

In order that this invention be more fully understood, the followingexamples are set forth. These examples are for the purpose ofillustration only and are not to be construed as limiting the scope ofthe invention in any way.

In the following Examples, proton NMR spectra were obtained at 300 MHz;chemical shifts are quoted downfield from internal tetramethylsilane.

EXAMPLE 1 Assay for Inhibition of ICE/ced-3 Protease Family Activity

A. Determination of IC₅₀ Values

Fluorescence enzyme assays detecting the activity of the compounds ofFormula I utilizing the recombinant ICE and CPP32 enzymes are performedessentially according to Thornberry et al. (Nature 356:768:774 (1992))and Nicholson et al. (Nature 376:37-43 (1995)) respectively, (hereinincorporated by reference) in 96 well microtiter plates. The substrateis Acetyl-Tyr-Val-Ala-Asp-amino-4-methylcoumarin (AMC) (SEQ ID NO:1) forthe ICE assay and Acetyl-Asp-Glu-Val-Asp-amino-4-methylcoumarin (SEQ IDNO:2) for the CPP32, Mch2, Mch3 and Mch5 assays. Enzyme reactions arerun in ICE buffer (25 mM HEPES, 1 mM EDTA, 0.1% CHAPS, 10% sucrose, pH7.5) containing 2 mM DTT at room temperature in duplicate. The assaysare performed by mixing the following components:

50 μL ICE, Mch2, Mch5, CPP32 (18.8, 38, 8.1 and 0.153 nM concentrations,respectively) or Mch3 (1 unit) enzyme in ICE buffer containing either8.0 (ICE, Mch2, Mch3, CPP32) or 20 (Mch5) mM DTT;

50 μL compound of Formula I or ICE buffer (control); and

100 μL of 20 μM substrate.

The enzyme and the compound of Formula I to be assayed are allowed topreincubate in the microtitre plate wells for 30 minutes at roomtemperature prior to the addition of substrate to initiate the reaction.Fluorescent AMC product formation is monitored for one hour at roomtemperature by measuring the fluorescence emission at 460 nm using anexcitation wavelength of 360 nm. The fluorescence change in duplicate(control) wells are averaged and the mean values are plotted as afunction of inhibitor concentration to determine the inhibitorconcentration producing 50% inhibition (IC₅₀).

B. Determination of the Dissociation Constant Ki and Irreversible RateConstant k₃ for Irreversible Inhibitors

For the irreversible inhibition of a ICE/ced-3 Family Protease enzymewith a competitive irreversible inhibitor; using the model representedby the following formulas:

The product formation at time t may be expressed as: $\begin{matrix}{\lbrack P\rbrack_{t} = {\lbrack E\rbrack^{T}\left( \frac{\lbrack S\rbrack K_{i}}{\lbrack I\rbrack K_{s}} \right){\left( \frac{k_{s}}{k_{3}} \right)\left\lbrack {1 - e^{{- k_{3}}{t/{({1 + {\frac{K_{i}}{\lbrack I\rbrack}{({1 + \frac{\lbrack S\rbrack}{K_{s}}})}}})}}}} \right\rbrack}}} & \text{Equation~~1}\end{matrix}$

where E, I, EI and E-I denote the active enzyme, inhibitor, non-covalentenzyme-inhibitor complex and covalent enzyme-inhibitor adduct,respectively. The K_(i) value is the overall dissociation constant ofthe reversible binding steps, and k₃ is the irreversible rate constant.The [S] and K_(s) values are the substrate concentration anddissociation constant of the substrate bound to the enzyme,respectively. [E]^(T) is the total enzyme concentration.

EXAMPLE 2 Synthesis of Representative Compound

Bromomethylketone (2):

4-Methylmorpholine (0.75 mL, 6.8 mmol) was added to a solution ofFmoc-Asp(OBn)-OH (1) (2.03 g, 4.55 mmol) in 50 mL of dry THF at −10° C.under an atmosphere of nitrogen, followed by the addition of isobutylchloroformate (0.78 mL, 6.0 mmol), and the solution was stirred for 20minutes. The resulting white precipitate was removed by filtration andthe filtrate was cooled to 0° C. In a separate flask,1-methyl-3-nitro-1-nitrosoguanidine (1.08 g, 7.36 mmol) was added to avigorously stirred mixture of diethyl ether (14 mL) and 40% KOH (8 mL)at 0° C. The resulting mixture was stirred for 10 minutes and the layerswere allowed to separate. The ether layer was transferred via plasticpipette to the original filtrate in THF and the reaction mixture wasstirred for 30 minutes. Then, 48% HBr in water (2.10 mL) was added andthe reaction mixture was warmed to room temperature over 15 minutes. Thesolution was diluted with ethyl acetate, washed twice with saturatedaqueous sodium bicarbonate, once with brine, dried (MgSO₄), andconcentrated. The resulting crude product was purified by flashchromatography on silica gel, eluting with 35% ethyl acetate-hexanes, toafford 1.73 g (73%) of (2) as a white solid. ¹H-NMR (300 MHz, CDCl₃):δ7.77 (d, J=8 Hz, 2H), 7.58 (d, J=8 Hz, 2H), 7.45-7.29 (m, 9H), 5.77 (d,J=9 Hz, 1H), 5.12 (s, 2H), 4.79-4.71 (m, 1H), 4.63-4.42 (m, 2H), 421 (t,j=6 Hz, 1H), 4.04 (s, 2H), 2.97 (ABXq, J=17, 5 Hz, 2H).

Ketone (3):

Sodium iodide (108 mg, 0.720 mmol) was added to a solution of 2 (1.72 g,3.28 mmol) in 10 mL of acetone at room temperature, followed by theaddition of the potassium salt of 2,3,5,6-tetrafluorophenol (704 mg,3.45 mmol) and the resulting mixture was stirred for one hour. Thereaction mixture was diluted with ethyl acetate, washed twice withbrine, dried (MgSO₄), and concentrated. The crude product was purifiedby flash chromatography on silica gel, eluting with 1:1:3dichloromethane/diethyl ether/hexanes, to provide 1.60 g (80%) of (3) asa white solid. ¹H-NMR (300 MHz, CDCl₃): δ7.76 (d, J=8 Hz, 2H), 7.58 (d,J=8Hz, 2H), 7.44-7.27 (m, 9H), 6.85-6.73 (m, 1H), 5.73 (d, J=9 Hz, 1H),5.15-4.92 (m, 4H), 4.75-4.67 (m, 1H), 4.61-4.42 (m, 2H), 4.21 (t, J=6Hz, 1H), 3.00 (ABXq, J=18, 4 Hz, 2H).

Alcohol (4):

Sodium borohydride (121 mg, 3.20 mmol) was added to a solution of 3(1.60 g, 2.63 mmol) in 7 mL of dry methanol and 7 mL of dry THF at 0° C.and the resulting mixture was stirred for 30 minutes. The reactionmixture was quenched with saturated aqueous ammonium chloride solution,extracted three times with dichloromethane, and the combineddichloromethane layers were washed once with brine, dried (MgSO₄), andconcentrated. The crude product was purified by flash chromatography onsilica gel, eluting with 50% ethyl acetate-hexanes, to give 1.39 g (87%)of (4) as a white solid. ¹H-NMR (300 MHz, CDCl₃): δ7.78-7.74 (m, 2H),7.57 (d, J=7 Hz, 2H), 7.44-7.27 (m, 9H), 6.87-6.75 (m, 1H), 5.62 (d, J=9Hz, 0.3H), 5.44 (d, J=9 Hz, 0.2H), 5.29-5.23 (m, 0.5H), 5.16-5.11 (m,1H), 4.69 (d, J=6 Hz, 1H), 4.59-4.37 (m, 4H), 4.30-4.04 (m, 3H),3.35-3.09 (m, 1H), 2.94-2.41 (m, 2H).

THP Ether (5):

3,4-Dihydro-2H-pyran (0.31 mL, 3.4 mmol) and pyridiniump-toluenesulfonate (111 mg, 0.441 mmol) were added to a solution of 4(1.39 g, 2.28 mmol) in 12 mL of dry dichloromethane and the resultingsolution was stirred at room temperature for 16 hours. The reactionmixture was diluted with ethyl acetate, washed twice with saturatedaqueous sodium bicarbonate solution, once with brine, dried (MgSO₄), andconcentrated. The crude product was purified by flash chromatography onsilica gel, first eluting with 15% ethyl acetate-hexanes and then with50% ethyl acetate-hexanes, to afford 1.09 g (69%) of (5) as a colorlessoil. ¹H-NMR (300 MHz, CDCl₃): δ7.76 (d, J=7 Hz, 2H), 7.62-7.55 (m, 2H),7.42-7.27 (m, 9H), 6.84-6.71 (m, 1H), 6.21 (d, J=9 Hz, 0.3H), 5.65 (d,J=9 Hz, 0.2H), 5.33-5.27 (m, 0.5H), 5.13 (t, J=3 Hz, 2H), 4.72-4.04 (m,8H), 3.91-3.73 (m, 1H), 3.51-3.36 (m, 1H), 2.98-2.57 (m, 2H), 1.86-1.61(m, 2H), 1.57-1.43 (m, 4H).

Amine (6):

Piperidine (0.50 mL, 5.1 mmol) was added to a solution of 5 (1.09 g,1.57 mmol) in 10 mL of dry DMF at room temperature and the resultingsolution was stirred for 5 minutes. The reaction mixture was dilutedwith ethyl acetate, washed once with saturated aqueous ammonium chloridesolution, twice with water, once with brine, dried (MgSO₄), andconcentrated. The crude product was purified by flash chromatography onsilica gel, first eluting with 50% ethyl acetate-hexanes and then with80% ethyl acetate-hexanes, to provide 544 mg (74%) of (6) as a colorlessoil. ¹H-NMR (300 MHz, CDCl₃): δ7.39-7.29 (m, 5H), 6.82-6.70 (m, 1H),5.15 (s, 2H), 4.78-4.63 (m, 1H), 4.53-4.26 (m, 2H), 4.03-3.79 (m, 2H),3.71-3.43 (m, 2H), 2.80-2.43 (m, 2H), 1.85-1.66 (m, 2H), 1.57-1.45 (m,4H).

Dipeptide (7):

Amine 6 (544 mg, 1.15 mmol) and Fmoc-Val-OH (433 mg, 1.28 mmol) weredissolved in 30 mL of dry dichloromethane. 1-Hydroxybenzotriazolehydrate (237 mg, 1.76 mmol) was added to this solution, followed by theaddition of 4-methylmorpholine (0.19 mL, 1.7 mmol) and1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (271 mg,1.41 mmol), and the resulting mixture was stirred at room temperaturefor 16 hours. The reaction mixture was diluted with ethyl acetate,washed once with saturated aqueous ammonium chloride solution, once withsaturated aqueous sodium bicarbonate solution, once with brine, dried(MgSO₄), and concentrated. The crude product was purified by flashchromatography on silica gel, eluting with 35% ethyl acetate-hexanes, togive 837 mg (91%) of (7) as a white solid. ¹H-NMR (300 MHz, CDCl₃):δ7.76 (d, J=8Hz, 2H), 7.62-7.56 (m, 2H), 7.42-7.27 (m, 9H), 6.91-6.71(m, 2H), 5.39-5.30 (m, 1H), 5.12-5.05 (m, 2H), 4.74-3.78 (m, 10H),3.50-3.36 (m, 1H), 2.97-2.61 (m, 2H), 2.19-2.06 (m, 1H), 1.82-1.68 (m,2H), 1.52-1.40 (m, 4H), 0.98-0.87 (m, 6H).

Amine (8):

Piperidine (0.35 mL, 3.6 mmol) was added to a solution of 7 (830 mg,1.05 mmol) in 7 mL of dry DMF at room temperature and the resultingsolution was stirred for 5 minutes. The reaction mixture was dilutedwith ethyl acetate, washed once with saturated aqueous ammonium chloridesolution, twice with water, once with brine, dried (MgSO₄), andconcentrated. The crude product was purified by flash chromatography onsilica gel, first eluting with 50% ethyl acetate-hexanes and then with20% methanol-dichloromethane, to provide 597 mg (100%) of(8) as a yellowoil. ¹H-NMR (300 MHz, CDCl₃): δ8.07-7.63 (m, 1H), 7.40-7.27 (m, 5H),6.83-6.71 (m, 1H), 5.17-5.07 (m, 2H), 4.76-4.62 (m, 1H), 4.60-4.26 (m,2H), 4.24-4.09 (m, 2H), 3.92-3.80 (m, 1H), 3.52-3.39 (m, 1H), 3.22-3.16(m, 1H), 2.97-2.60 (m, 2H), 2.31-2.16 (m, 1H), 1.84-1.64 (m, 2H),1.59-1.44 (m, 4H), 0.98-0.94 (m, 3H), 0.81-0.76 (m, 3H).

Dipeptide (9):

Amine 8 (578 mg, 1.01 mmol) and (1-naphthoxy)acetic acid (226 mg, 1.12mmol) were dissolved in 25 mL of dry dichloromethane.1-Hydroxybenzotriazole hydrate (211 mg, 1.56 mmol) was added to thissolution, followed by the addition of N-methylmorpholine (0.17 mL, 1.5mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride(239 mg, 1.25 mmol), and the resulting mixture was stirred at roomtemperature for 16 hours. The reaction mixture was diluted with ethylacetate, washed once with saturated aqueous ammonium chloride solution,once with saturated aqueous sodium bicarbonate solution, once withbrine, dried (MgSO₄), and concentrated. The crude product was purifiedby flash chromatography on silica gel, eluting with 35% ethylacetate-hexanes, to give 682 mg (89%) of (9) as a white foam. ¹H-NMR(300 MHz, CDCl₃): δ8.28-8.21 (m, 1H), 7.85-7.80 (m, 1H), 7.57-7.49 (m,3H), 7.40-7.22 (m, 8H), 6.96-6.42 (m, 2H), 5.12-5.05 (m, 2H), 4.80-4.59(m, 3H), 4.58-4.24 (m, 3H), 4.23-4.06 (m, 2H), 3.96-3.77 (m, 1H),3.56-3.38 (m, 1H), 2.97-2.61 (m, 2H), 2.24-2.10 (m, 1H), 1.84-1.66 (m,2H), 1.55-1.40 (m, 4H), 0.98-0.87 (m, 6H).

Acid (10):

10% Palladium on carbon (170 mg) was added to a solution of 9 (650 mg,0.861 mmol) in anhydrous methanol (15 mL) under an atmosphere ofnitrogen and the flask was then evacuated with the house vacuum. Themixture was stirred under a balloon of hydrogen gas for 75 minutes, thenfiltered through Celite, and eluted with methanol. The solution wasconcentrated to afford 345 mg (94%) of (10) as a white solid.

Methyl sulfonimide (11):

1,1′-Carbonyldiimidazole (146 mg, 0.900 mmol) was added to a solution of10 (300 mg, 0.451 mmol) in dry THF (7 mL) under an atmosphere ofnitrogen, and the reaction mixture was stirred for 3 hours. The mixturewas cooled to 0° C., and the methanesulfonamide (86 mg, 0.90 mmol) wasadded, followed by the addition of 1,8-diazabicyclo[5.4.0]undec-7-ene(0.135 mL, 0.903 mmol). The resulting mixture was stirred at roomtemperature for 4 hours. The reaction mixture was diluted with ethylacetate, washed once with 1 N HCl solution, twice with water, once withbrine, dried (MgSO₄), and concentrated. The residue was reconcentratedfrom dichloromethane to provide 314 mg (98%) of (1) as a white solid.¹H-NMR (300 MHz, CDCl₃): δ10.22-9.99 (m, 1H), 8.24-8.17 (m, 1H),7.85-7.69 (m, 1H), 7.59-7.47 (m, 3H), 7.40-7.27 (m, 2H), 6.94-6.66 (m,2H), 4.85-4.62 (m, 4H), 4.55-4.02 (m, 6H), 3.59-3.45 (m, 1H), 3.22-3.19(m, 3H), 2.89-2.55 (m, 2H), 2.21-2.09 (m, 1H), 1.88-1.67 (m, 2H),1.63-1.43 (m, 4H), 1.00-0.86 (m, 6H).

Alcohol (12):

p-Toluenesulfonic acid (34 mg, 0.18 mmol) was added to a solution of 11(302 mg, 0.426 mmol) in anhydrous methanol (5 mL) and the reactionmixture was stirred at room temperature for 30 minutes. The mixture wasdiluted with ethyl acetate, washed twice with water, once with brine,dried (MgSO₄), and concentrated to afford 249 mg (93%) of (12) as awhite solid. ¹H-NMR (300 MHz, DMSO): δ8.23-8.18 (m, 2H), 8.04-7.88 (m,2H), 7.61-7.50 (m, 4H), 7.43-7.37 (m, 1H), 6.90 (d, J=8 Hz, 1H), 5.61(d, J=5 Hz, 0.3H), 5.48 (d, J=6 Hz, 0.7H), 4.83-4.72 (m, 2H), 4.58-3.75(m, 5H), 3.17 (s, 1H), 3.13 (s, 2H), 2.74-2.37 (m, 2H), 2.03-1.91 (m,1H), 0.84-0.76 (m, 6H).

Methylsulfonimide (“Compound No. 1”):

Dess-Martin periodinane (203 mg, 0.479 mmol) was added to a solution of12 (233 mg, 0.373 mmol) in 7 mL of dry dichloromethane, and the reactionmixture was stirred at room temperature for 30 minutes. The mixture wasdiluted with ethyl acetate, washed twice with water, once with brine,dried (MgSO₄), and concentrated. The crude product was purified by flashchromatography, eluting first with 60% ethyl acetate-hexanes and thenwith 80% ethyl acetate-hexanes, to provide 134 mg (58%) of Compound No.1 as a mixture of diastereomers and as a white solid. ¹H-NMR (300 MHz,DMSO): δ8.84 (d, J=7 Hz, ¹H), 8.28-8.06 (m, 2H), 7.91-7.87 (m, 1H),7.58-7.49 (m, 5H), 7.39 (t, J=8 Hz, 1H), 6.89 (d, J=8 Hz, 1H), 5.23 (s,1H), 4.93-4.42 (m, 4H), 4.28-4.19 (m, 1H), 3.28 (s, 1.3H), 3.19 (s,1.7H), 2.96-2.59 (m, 2H), 2.06-1.96 (m, 1H), 0.89-0.80 (m, 6H); MS (ESI)m/e 654 [(M+)−1].

EXAMPLE 3 Alternative Synthesis of Compounds of Formula I

This example illustrates synthesis of compounds of Formula I byformation of a stabilized sulfonamide ring, followed by addition viaamide bond formation to the remainder of the compound. In the followingrepresentative examples, q is 1, r is 2 and R is methyl.

Example Scheme 3a

Starting with the commercially available Z-Asp(OtBu)-OH, the Weinrebamide is formed, followed by hydrolysis of the t-butyl ester. The betacarboxylic acid is then coupled with methyl sulfonamide (or othersubstituted sulfonamide), followed by reduction of the Weinreb amide tothe aldehyde. Acid-catalyzed acetal formation using ethanol is assistedby cyclization of the sulfonamide to form a stable 5-membered ring. Thecarbobenzyloxy urethane is then removed, the aspartyl intermediatecoupled to the substituted acyl peptide of choice, then the acetaldeprotected. (a) EDCl, HOBt, N-Me Morpholine, HClH₂N(OCH₃)CH₃, CH₂Cl₂,0° C.-RT; (b) TFA, anisole, CH₂Cl₂, RT; (c) EDCl, DMAP (cat.),CH₃SO₂NH₂, CH₂Cl₂, RT; or i. CDI; ii. CH₃SO₂NH₂, DBU, 0° C. (d) LAH,THF, 0° C.; (e) Ethanol, CH(OEt)₃, p-TsOH, toluene reflux; (f) H₂; Pd/C,RT; (g) R¹-X-(CH₂)_(n)—CH(R²)CO(amino acid)CO₂H, EDCl, HOBt, N-MeMorpholine, CH₂Cl₂, 0° C.-RT; (h) TFA, anisole, CH₂Cl₂, H₂O RT.

Example Scheme 3b

Starting with the commercially available Fmoc-Asp(OtBu)-OH, the Weinrebamide is formed, followed by hydrolysis of the t-butyl ester. The betacarboxylic acid is then coupled with methyl sulfonamide (or othersubstituted sulfonamide), followed by reduction of the Weinreb amide tothe aldehyde. Acid-catalyzed acetal formation using benzyl alcohol isassisted by cyclization of the sulfonamide to form a stable 5-memberedring. The fluorenylmethyloxy urethane is then removed, the aspartylintermediate coupled to the substituted acyl peptide of choice, then theacetal deprotected. (a) EDCl, HOBt, N-Me Morpholine, HClH₂N(OCH₃)CH₃,CH₂Cl₂, 0° C.-RT; (b) TFA, anisole, CH₂Cl₂, RT; (c) EDCI, DMAP (cat.),CH₃SO₂NH₂, CH₂Cl₂, RT; (d) LAH, THF, 0° C.; (e) Benzyl alcohol, p-TsOH,toluene reflux; (f) Et₂NH, DMF, RT; (g) R¹-X-(CH₂)_(n)—CH(R²)CO(aminoacid)CO₂H, EDCl, HOBt, N-Me Morpholine, CH₂Cl₂, 0° C.-RT; (h) H₂; Pd/C,RT.

Example Scheme 3c

Starting from the readily available intermediate, modified by additionof tetrafluorophenyl, the t-butyl ester is hydrolyzed, followed bycoupling with methyl sulfonamide (or other substituted sulfonamide).Acid-catalyzed ketal formation using ethanol is assisted by cyclizationof the sulfonamide to form a stable 5-membered ring. The carbobenzyloxyurethane is then removed, the aspartyl intermediate coupled to thesubstituted acyl peptide of choice, then the ketal deprotected. (a) TFA,anisole, CH₂Cl₂, RT; (b) EDCl, DMAP (cat.), CH₃SO₂NH₂, CH₂Cl₂, RT; or i.CDI; ii. CH₃SO₂NH₂, DBU, 0° C.; (c) Ethanol, CH(OEt)₃, p-TsOH, toluenereflux; (d) H₂; Pd/C, RT; (e) R¹-X(CH₂)_(n)—CH(R²)CO(amino acid)CO₂H,EDCl, HOBt, N-Me Morpholine, CH₂Cl₂, 0° C.-RT; (f) TFA, anisole, CH₂Cl₂,H₂O, RT.

Example Scheme 3d

Starting from the readily available intermediate, modified by additionof tetrafluorophenyl, the t-butyl ester is hydrolyzed, followed bycoupling with methyl sulfonamide (or other substituted sulfonamide).Acid-catalyzed ketal formation using ethanol is assisted by cyclizationof the sulfonamide to form a stable 5-membered ring. Thefluorenylmethyloxy urethane is then removed, the aspartyl intermediatecoupled to the substituted acyl peptide of choice, then the ketaldeprotected. (a) TFA, anisole, CH₂Cl₂, RT; (b) EDCl, DMAP (cat.),CH₃SO₂NH₂, CH₂Cl₂, RT; (c) Ethanol, CH(OEt)₃, p-TsOH, toluene reflux;(d) Et₂NH, DMF, RT; (e) R¹-X-(CH₂)_(n)—CH(R²)CO(amino acid)CO₂H, EDCl,HOBt, N-Me Morpholine, CH₂Cl₂, 0° C.-RT; (f) H₂; Pd/C, RT.

EXAMPLE 4 Representative Compounds

The representative compounds listed in the following Table 1 may be madeaccording to the procedures set forth in Examples 2 and 3.

TABLE 1 Representative Compounds

No. A B R¹ 2 NHCH(CH₂CH(CH₃)₂)CO CH₂F 1-naphthyl 3 NHCH(CH(CH₃)₂)CO CH₂F1-naphthyl 4 NHCH(CH(CH₃)₂)CO CH₂OCO(2,4-diCl—Ph) 1-naphthyl 5NHCH(CH(CH₃)₂)CO CH₂O(2,6-diF—Ph) 1-naphthyl 6 NHCH(CH(CH₃)₂)COCH₂O(2,4,6-triF—Ph) 1-naphthyl 7 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) 1-naphthyl 8 NHCH(CH(CH₃)₂)COCH₂O(6-Me-2-pyron-4-yl) 1-naphthyl 9 NHCH(CH(CH₃)₂)COCH₂O(2-Ph-5,6-benzopyran-4- 1-naphthyl on-3-yl) 10 NHCH(CH(CH₃)₂)COCH₂OPO(Me)Ph 1-naphthyl 11 NHCH(CH(CH₃)₂)CO CH₂OPOPh₂ 1-naphthyl 12NHCH(CH(CH₃)₂)CO CH₂O(2-CF₃-pyrimidin-4-yl) 1-naphthyl 13NHCH(CH(CH₃)₂)CO CH₂O(5-CO₂Me-isoxazol-3-yl) 1-naphthyl 14NHCH(CH(CH₃)₂)CO CH₂OPO(Me)(1-naphthyl) 1-naphthyl 15NHCH(CH₂CH(CH₃)₂)CO CH₂OPOPh₂ 1-naphthyl 16 NHCH(CH₂CH(CH₃)₂)COCH₂OCO(2,6-diCl—Ph) 1-naphthyl 17 NHCH(CH₂CH(CH₃)₂)COCH₂O(2,4,6-triF—Ph) 1-naphthyl 18 NHCH(CH₂CH(CH₃)₂)CO CH₂O(2,3,5,6-tetraF—Ph) 1-naphthyl 19 NHCH(CH₂CH(CH₃)₂)CO CH₂OPO(Me)Ph1-naphthyl 20 NHCH(CH₃)CO CH₂O(2-F—Ph) (2-Ph)Ph 21 NHCH(CH₃)COCH₂OCO(2,6-di-Cl—Ph) (2-Ph)Ph 22 NHCH(CH₃)CO CH₂OPOPh₂ (2-Ph)Ph 23NHCH(CH₃)CO CH₂O(2-F—Ph) (2-t-Bu)Ph 24 NHCH(CH₃)CO CH₂OPOPh₂ (2-t-Bu)Ph25 NHCH(CH₃)CO CH₂OCO(2,3,5,6-tetra-Cl—Ph) 1-naphthyl-CH₂ 26 NHCH(CH₃)COCH₂OCO(2,6-di-Cl—Ph) 1-naphthyl-CH₂ 27 NHCH(CH₃)CO CH₂OPOPh₂1-naphthyl-CH₂ 28 NHCH(CH(CH₃)₂)CO CH₂O(2,3,5,6-tetraF—Ph)1-naphthyl-CH₂ 29 NHCH(CH(CH₃)₂)CO CH₂O(2,3,5,6-tetraF—Ph) PhCH₂ 30NHCH(CH(CH₃)₂)CO CH₂O(2,3,5,6-tetraF—Ph) Ph(CH₂)₂ 31 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) Ph₂CH 32 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) Ph 33 NHCH(CH(CH₃)₂)CO CH₂O(2,3,5,6-tetraF—Ph)(2-Ph)Ph 34 NHCH(CH(CH₃)₂)CO CH₂O(2,3,5,6-tetraF—Ph) (2-PhCH₂)Ph 35NHCH(CH(CH₃)₂)CO CH₂O(2,3,5,6-tetraF—Ph) (3-PhO)Ph 36 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) 4-Cl-1-naphthyl 37 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) 2-anthryl 38 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) 2-benzimidazolyl 39 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) 1-adamantanyl 40 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) (2-F)Ph 41 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) (4-F)Ph 42 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) (2-CF₃)Ph 43 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) (2-t-Bu)Ph 44 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) (4-n-heptyl)Ph 45 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) (2-CH₃O)Ph 46 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) (2-PhO)Ph 47 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) 2-naphthyl 48 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) 5,6,7,8-tetrahydro-1- naphthyl 49NHCH(CH(CH₃)₂)CO CH₂O(2,3,5,6-tetraF—Ph) 1-anthryl 50 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) 2-pyridinyl 51 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) 4-pyridinyl 52 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) 2,3,5,6-tetrafluoro-4- pyridinyl 53NHCH(CH(CH₃)₂)CO CH₂O(2,3,5,6-tetraF—Ph) 2-pyrazinyl 54 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) 1,2,3,4-tetrahydro-1- naphthyl 55NHCH(CH(CH₃)₂)CO CH₂O(2,3,5,6-tetraF—Ph) (2-Cl)Ph 56 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) (2-Br)Ph 57 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) (2-I)Ph 58 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) (2,6-di-F)Ph 59 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) (2,5-di-t-Bu)Ph 60 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) 5-indanyl 61 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) (3,4,5-tri-MeO)PhCH₂ 62 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) methyl 63 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) n-heptyl 64 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) t-octyl 65 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) cyclo-hexyl 66 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) 5-Ph-3-pyrazolyl 67 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) (2-F-4-I)Ph 68 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) (2,3,4,5-tetra-F)Ph 69 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) (2,3,4,6-tetra-F)Ph 70 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) (2,3,5,6-tetra-Cl)Ph 71 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) (2,3,4,5,6-penta-F)Ph 72 NHCH(CH(CH₃)₂)COCH₂O(2,3,5,6-tetraF—Ph) Ph₂N 73 NHCH(CH(CH₃)₂)CO CH₂O(2,3,5,6-tetraF—Ph)PHCH₂(Ph)N 74 NHCH(CH(CH₃)₂)CO CH₂O(2,3,5,6-tetraF—Ph) PhCH₂O 75NHCH(CH₃)CO CH₂O(2,3,5,6-tetraF—Ph) (2-t-Bu)Ph 76 NHCH(CH₃)COCH₂O(2,3,5,6-tetraF—Ph) (2-CF₃)Ph 77 NHCH(CH₃)CO CH₂O(2,3,5,6-tetraF—Ph)(2-Ph)Ph 78 NHCH(CH₃)CO CH₂O(2,3,5,6-tetraF—Ph) (2-PhCH₂)Ph 79NHCH(CH₃)CO CH₂O(2,3,5,6-tetraF—Ph) (2-PhO)Ph 80 NHCH(CH₃)COCH₂O(2,3,5,6-tetraF—Ph) (3-PhO)Ph 81 NHCH(CH₃)CO CH₂O(2,3,5,6-tetraF—Ph)5,6,7,8-tetrahydro-1- naphthyl 82 NHCH(CH₃)CO CH₂O(2,3,5,6-tetraF—Ph)1-naphthyl 83 NHCH(CH₃)CO CH₂O(2,3,5,6-tetraF—Ph) Ph 84 NHCH(CH₃)COCH₂O(2,3,5,6-tetraF—Ph) (2,6-di-F)Ph 85 NHCH(CH₃)COCH₂O(2,3,5,6-tetraF—Ph) (4-Ph)Ph 86 NHCH(CH₃)CO CH₂O(2,3,5,6-tetraF—Ph)(4-MeO)Ph 87 NHCH(CH₃)CO CH₂O(2,3,5,6-tetraF—Ph) Ph₂CH 88NHCH(CH₂cyclohexyl)CO CH₂O(2,3,5,6-tetraF—Ph) (2-PhO)Ph 89NHCH(CH₂cyclohexyl)CO CH₂O(2,3,5,6-tetraF—Ph) (2-Ph)Ph 90NHCH(CH₂cyclohexyl)CO CH₂O(2,3,5,6-tetraF—Ph) (2-PhCH₂)Ph 91NHCH(CH₂cyclohexyl)CO CH₂O(2,3,5,6-tetraF—Ph) 1-naphthyl 92NHCH(CH₂cyclohexyl)CO CH₂OCO(2,6-diCl—Ph) 5,6,7,8-tetrahydro-1- naphthyl93 NHCH(CH₂cyclohexyl)CO CH₂O(2,3,5,6-tetra-F—Ph) 5,6,7,8-tetrahydro-1-naphthyl 94 NHCH(CH₂cyclohexyl)CO CH₂OPO(Me)Ph 5,6,7,8-tetrahydro-1-naphthyl 95 NHCH(CH₂cyclohexyl)CO CH₂OPOPh₂ 5,6,7,8-tetrahydro-1-naphthyl 96 NHCH(CH₂cyclohexyl)CO CH₂OPO(Me)Ph (2-PhCH₂)Ph 97NHCH(CH₂cyclohexyl)CO CH₂OPOPh₂ (2-PhCH₂)Ph 98 NHCH(CH₂cyclohexyl)COCH₂OPO(Me)Ph (2-Ph)Ph 99 NHCH(CH₂cyclohexyl)CO CH₂OPOPh₂ (2-Ph)Ph 100

CH₂O(2,3,5,6-tetra-F—Ph) 1-naphthyl 101

CH₂O(2,3,5,6-tetra-F—Ph) 1-naphthyl 102 NHCH(cyclohexyl)COCH₂O(2,3,5,6-tetra-F—Ph) 1-naphthyl 103 NorleucineCH₂O(2,3,5,6-tetra-F—Ph) 1-naphthyl 104 (t-butyl)glycineCH₂O(2,3,5,6-tetra-F—Ph) 1-naphthyl 105 (t-butyl)alanineCH₂O(2,3,5,6-tetra-F—Ph) 1-naphthyl 106 PhenylglycineCH₂O(2,3,5,6-tetra-F—Ph) 1-naphthyl 107 PhenylalanineCH₂O(2,3,5,6-tetra-F—Ph) 1-naphthyl 108 HomophenylalanineCH₂O(2,3,5,6-tetra-F—Ph) 1-naphthyl 109 1-aminocyclopentaneCH₂O(2,3,5,6-tetra-F—Ph) 1-naphthyl carboxylic acid 110NHCH(CH₂CH₂SOCH₃)CO CH₂O(2,3,5,6-tetra-F—Ph) 1-naphthyl 111

H 1-naphthyl 112 NHCH(CH(CH₃)₂)CO H 2-(1H-tetrazol-5-yl)Ph 113NHCH(CH(CH₃)₂)CO H 1-adamantanyl 114 NHCH(CH(CH₃)₂)CO H Ph 115NHCH(CH(CH₃)₂)CO H PhCH₂ 116 NHCH(CH(CH₃)₂)CO H Ph(CH₂)₂ 117NHCH(CH(CH₃)₂)CO H (2-CF₃)Ph 118 NHCH(CH(CH₃)₂)CO H (2-t-Bu)Ph 119NHCH(CH(CH₃)₂)CO H (2-Ph)Ph 120 NHCH(CH(CH₃)₂)CO H (2-PhCH₂)Ph 121NHCH(CH(CH₃)₂)CO H (2-PhO)Ph 122 NHCH(CH(CH₃)₂)CO H 2-naphthyl 123NHCH(CH(CH₃)₂)CO H 1-naphthyl 124 NHCH(CH(CH₃)₂)CO H 4-Cl-1-naphthyl 125NHCH(CH(CH₃)₂)CO H 5,6,7,8-tetrahydro-1- naphthyl 126 NHCH(CH(CH₃)₂)CO H1,2,3,4-tetrahydro-1- naphthyl 127 NHCH(CH(CH₃)₂)CO H (1-naphthyl)CH₂128 NHCH(CH₂CH(CH₃)₂)CO H 1-naphthyl

EXAMPLE 5 Activity of Representative Compound

The activity of a representative compound of this invention (i.e.,Compound No. 1) was evaluated according to the procedures disclosed inExample 1. More specifically, the IC₅₀ and K_(i) for Compound No. 1 weredetermined as set forth above. The IC₅₀ results are provided in Table 2,as run against Cbz-ValAlaAsp-H as a reference control.

TABLE 2 Csp-1 Csp-3 Csp-6 Csp-7 Csp-8 Csp-9 IC₅₀ IC₅₀ IC₅₀ IC₅₀ IC₅₀IC₅₀ Cpd. No. (μM) (μM) (μM) (μM) (μM) (μM) 1 0.004 0.002 0.002 0.0040.006 0.005 Reference 0.064 47.0 >10 >10 2.96 0.87

The equations set forth in Example 1 were also used determine the K_(i)values of inhibitor (i.e., Compound No. 1) bound to a ICE/ced-3 familyprotease. Thus, a continuous assay was run for sixty minutes at variousconcentrations of the inhibitor and the substrate. The assay wasformulated essentially the same as described above for generating thedata in Table 2, except that the reaction was initiated by adding theenzyme to the substrate-inhibitor mixture. The K_(i) values wereobtained by simulating the product AMC formation as a function of timeaccording to Equation 1. The results of this second assay are set forthbelow in Table 3, wherein the reference compound was Cbz-ValAlaAsp-CH₂F.

TABLE 3 Csp-1 Csp-3 Csp-6 Csp-8 Cpd. No. K_(i)(μM) K_(i)(μM) K_(i)(μM)K_(i)(μM) 1 0.20 0.08 0.40 0.60 reference 0.015 0.820 0.594 0.018

Although the invention has been described with reference to the examplesprovided above, it should be understood that various modifications canbe made without departing from the spirit of the invention. Accordingly,the invention is limited only by the claims.

2 1 4 PRT Artificial Sequence Substrate for the ICE assay. 1 Tyr Val AlaAsp 1 2 4 PRT Artificial Sequence Substrate for the CPP32, Mch2, Mch3and Mch5 assays. 2 Asp Glu Val Asp 1

We claim:
 1. A compound of the following formula:

wherein: n is 0, 1 or 2; q is 1 or 2; r is 1 or 2; R is lower alkyl,alkyl, cycloalkyl, (cycloalkyl)alkyl, phenyl, substituted phenyl,phenylalkyl, substituted phenylalkyl, naphthyl, substituted naphthyl, (1or 2 naphthyl)alkyl, substituted (1 or 2 naphthyl)alkyl, heteroaryl,substituted heteroaryl, (heteroaryl)alkyl, substituted(heteroaryl)alkyl, NR^(a)(R^(b)) or OR^(c); R¹ is phenyl, substitutedphenyl, naphthyl, substituted naphthyl, heteroaryl, or substitutedheteroaryl; R² is hydrogen, alkyl, cycloalkyl, phenyl, substitutedphenyl, (CH₂)_(p)CO₂R³, (CH₂)_(m)NH₂, (CH₂)_(m)NHCOR¹⁰,(CH₂)_(m)N(C═NH)NH₂, (CH₂)_(p)OR¹¹, (CH₂)_(p)SR¹², (CH₂)_(m)cycloalkyl,(CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or2-naphthyl), or (CH₂)_(m)heteroaryl; X is CH₂, C═O, O, S or NH; A is anatural or unnatural amino acid of Formula IIa-i:

 B is a hydrogen atom, a deuterium atom, C₁₋₁₀ straight chain orbranched alkyl, cycloalkyl, phenyl, substituted phenyl, naphthyl,substituted naphthyl, 2-benzoxazolyl, substituted 2-oxazolyl,(CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl),(CH₂)_(m)(1 or 2-naphthyl), (CH₂)_(m)heteroaryl, halomethyl, CO₂R¹³,CONR¹⁴R¹⁵, CH₂ZR¹⁶, CH₂OCO(aryl), CH₂OCO(substituted aryl),CH₂OCO(heteroaryl), CH₂OCO(substituted heteroaryl), or CH₂OPO(R¹⁷)R¹⁸,where Z is an oxygen or a sulfur atom, or B is a group of the FormulaIIIa-c:

 and wherein R^(a) and R^(b) are the same or different and independentlyhydrogen, alkyl, cycloalkyl, (cycloalkyl)alkyl, phenyl, substitutedphenyl, phenylalkyl, substituted phenylalkyl, naphthyl, substitutednaphthyl, (1 or 2 naphthyl)alkyl, substituted (1 or 2 naphthyl)alkyl,heteroaryl, substituted heteroaryl, (heteroaryl)alkyl, or substituted(heteroaryl)alkyl, with the proviso that R^(a) and R^(b) cannot both behydrogen; R^(c) is alkyl, cycloalkyl, (cycloalkyl)alkyl, phenyl,substituted phenyl, phenylalkyl, substituted phenylalkyl, naphthyl,substituted naphthyl, (1 or 2 naphthyl)alkyl, substituted (1 or 2naphthyl)alkyl, heteroaryl, substituted heteroaryl, (heteroaryl)alkyl,or substituted (heteroaryl)alkyl; R³ is hydrogen, alkyl, cycloalkyl,(cycloalkyl)alkyl, phenylalkyl, or substituted phenylalkyl; R⁴ is alkyl,cycloalkyl, phenyl, substituted phenyl, (CH₂)_(m)NH₂, (CH₂)_(m)NHCOR¹⁰,(CH₂)_(m)N(C═NH)NH₂, (CH₂)_(p)CO₂R³, (CH₂)_(p)OR¹¹, (CH₂)_(p)SR¹²,(CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl),(CH₂)_(m)(1 or 2-naphthyl), or (CH₂)_(m)heteroaryl; R^(4a) is hydrogen,or methyl; or R⁴ and R^(4a) taken together are —(CH₂)_(d)— where d is aninteger from 2 to 6; R⁵ is phenyl, substituted phenyl, (CH₂)_(p)phenyl,(CH₂)_(p)(substituted phenyl), cycloalkyl, or benzofused cycloalkyl; R⁶is hydrogen, alkyl, cycloalkyl, phenyl, substituted phenyl,(CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), or(CH₂)_(m)(1 or 2-naphthyl); R⁷ is hydrogen, fluorine, oxo, alkyl,cycloalkyl, phenyl, substituted phenyl, naphthyl, (CH₂)_(m)cycloalkyl,(CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or2-naphthyl), OR¹¹, SR¹², or NHCOR¹⁰; R⁸ is hydrogen, oxo, alkyl,cycloalkyl, phenyl, substituted phenyl, naphthyl, (CH₂)_(m)cycloalkyl,(CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), or (CH₂)_(m)(1 or2-naphthyl); R⁹ is alkyl, cycloalkyl, (CH₂)_(m)cycloalkyl,(CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or2-naphthyl), or COR¹⁰; R¹⁰ is hydrogen, alkyl, cycloalkyl, phenyl,substituted phenyl, naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or 2-naphthyl), OR¹³, orNR¹⁴R¹⁵; R¹¹ is hydrogen, alkyl, cycloalkyl, phenyl, substituted phenyl,naphthyl, (CH₂)_(m)cycloalkyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substitutedphenyl), or (CH₂)_(m)(1 or 2-naphthyl); R¹² is alkyl, cycloalkyl,phenyl, substituted phenyl, naphthyl, (CH₂)_(m)cycloalkyl,(CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), or (CH₂)_(m)(1 or2-naphthyl); R¹³ is alkyl, cycloalkyl, (CH₂)_(m)cycloalkyl,(CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), or (CH₂)_(m)(1 or2-naphthyl); R¹⁴ is hydrogen, alkyl, cycloalkyl, phenyl, substitutedphenyl, naphthyl, substituted naphthyl, (CH₂)_(m)cycloalkyl,(CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), or (CH₂)_(m)(1 or2-naphthyl); R¹⁵ is hydrogen or alkyl; or R¹⁴ and R¹⁵ taken togetherform a five, six or seven membered carbocyclic or heterocyclic ring,such as morpholine or N-substituted piperazine; R¹⁶ is phenyl,substituted phenyl, naphthyl, substituted naphthyl, heteroaryl,(CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1 or2-naphthyl), or (CH₂)_(m)heteroaryl; R¹⁷ and R¹⁸ are independentlyalkyl, cycloalkyl, phenyl, substituted phenyl, naphthyl, or phenylalkyl,substituted phenylalkyl, or (cycloalkyl)alkyl; R¹⁹ and R²⁰ areindependently hydrogen, alkyl, phenyl, substituted phenyl,(CH₂)_(m)phenyl, or (CH₂)_(m)(substituted phenyl), or R¹⁹ and R²⁰ takentogether are —(CH═CH)₂—; R²¹ is hydrogen, alkyl, phenyl, substitutedphenyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl); R²², R²³ and R²⁴are independently hydrogen or alkyl; Y¹ is CH₂, (CH₂)₂, (CH₂)₃, or S; Y²is O or NR²⁴; Y³ is CH₂, O, or NR²⁴; a is 0 or 1 and b is 1 or 2,provided that when a is 1 then b is 1; c is 1 or 2, provided that when cis 1 then a is 0 and b is 1; m is 1, 2, 3 or 4; and p is 1 or 2; or apharmaceutically acceptable salt thereof.
 2. The compound of claim 1where X is oxygen.
 3. The compound of claim 1 where X is sulfur.
 4. Thecompound of claim 1 where X is NH.
 5. The compound of claim 1 where X isCH₂.
 6. The compound of claim 1 where X is C═O.
 7. The compound of claim1 wherein q is
 1. 8. The compound of claim 1 wherein q is
 2. 9. Thecompound of claim 1 wherein A is


10. The compound of claim 9 wherein R⁴ is lower alkyl, cycloalkyl,phenyl, substituted phenyl, (CH₂)_(n)NH₂, (CH₂)_(m)OR¹⁰, (CH₂)_(m)SR¹¹,(CH₂)_(n)cycloalkyl, (CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl), or(CH₂)_(n)(1 or 2-naphthyl); and R^(4a) is hydrogen.
 11. The compound ofclaim 1 wherein A is


12. The compound of claim 11 wherein R⁵ is phenyl, substituted phenyl,(CH₂)_(m)phenyl, (CH₂)_(m)(substituted phenyl), cycloalkyl, or2-indanyl.
 13. The compound of claim 1 wherein A is


14. The compound of claim 13 wherein R⁷ is hydrogen, fluorine,cycloalkyl, phenyl, substituted phenyl, naphthyl, (CH₂)_(n)cycloalkyl,(CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl), (CH₂)_(n)(1 or2-naphthyl), OR¹⁰, or SR¹¹.
 15. The compound of claim 1 wherein A is


16. The compound of claim 15 wherein R⁸ is hydrogen, oxo, cycloalkyl,phenyl, substituted phenyl, or naphthyl; and Y¹ is CH₂, (CH₂)₂, (CH₂)₃,or S.
 17. The compound of claim 1 wherein A is


18. The compound of claim 17 wherein a is
 0. 19. The compound of claim 1wherein B is hydrogen, 2-benzoxazolyl, substituted 2-oxazolyl, CH₂ZR¹⁵,CH₂OCO(aryl), or CH₂OPO(R¹⁶)R¹⁷; and Z is O or S.
 20. The compound ofclaim 1 wherein B is


21. The compound of claim 20 wherein R¹⁹ and R²⁰ are independentlyhydrogen, alkyl, or phenyl, or wherein R¹⁹ and R²⁰ taken together are—(CH═CH)₂—.
 22. The compound of claim 1 wherein X is O or NH; n is 0 or1; q is 1; R is methyl; R¹ is substituted phenyl, naphthyl, orsubstituted naphthyl; and R² is hydrogen, lower alkyl, (CH₂)_(p)CO₂R³,(CH₂)_(m)(substituted phenyl), (CH₂)_(m)(1- or 2-naphthyl), or(CH₂)_(m)tetrazolyl.
 23. The compound of claim 22 wherein R¹ is1-naphthyl.
 24. The compound of claim 22 wherein R¹ is 2-naphthyl. 25.The compound of claim 22 wherein R¹ is substituted naphthyl.
 26. Thecompound of claim 25 wherein substituted naphthyl is2-carboxy-1-naphthyl.
 27. The compound of claim 22 wherein R¹ issubstituted phenyl.
 28. The compound of claim 27 wherein substitutedphenyl is 2-substituted phenyl.
 29. The compound of claim 28 wherein2-substituted phenyl is (2-phenyl)phenyl.
 30. The compound of claim 22wherein A is alanine, valine, leucine cyclohexylalanine, phenylgycine ort-butylglycine.
 31. The compound of claim 1 wherein R¹ is 1-naphthyl or2-naphthyl and A is valine.
 32. The compound of claim 1 wherein B isCH₂O(2,3,5,6-tetrafluorophenyl).
 33. The compound of claim 1 wherein R¹is substituted naphthyl.
 34. The compound of claim 1 wherein substitutednaphthyl is 2-carboxy-1-naphthyl.
 35. The compound of claim 1 wherein R¹is 2-substituted phenyl.
 36. The compound of claim 35 wherein2-substituted phenyl is (2-2phenyl)phenyl.
 37. The compound of claim 22wherein R² is (CH₂)₂CO₂R³ and n is
 0. 38. The compound of claim 22wherein R² is (CH₂)_(m)tetrazolyl and m is
 0. 39. The compound of claim1 wherein R is lower alkyl.
 40. The compound of claim 39 wherein R ismethyl.
 41. The compound of claim 1 wherein r is
 2. 42. The compound ofclaim 1 wherein r is
 1. 43. A composition comprising a compound of claim1 in combination with a pharmaceutically acceptable carrier.